110 results on '"Teboul L"'
Search Results
2. Genome wide conditional mouse knockout resources
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Kaloff, C, Anastassiadis, K, Ayadi, A, Baldock, R, Beig, J, Birling, M-C, Bradley, A, Brown, SDM, Bürger, A, Bushell, W, Chiani, F, Collins, FS, Doe, B, Eppig, JT, Finnell, RH, Fletcher, C, Flicek, P, Fray, M, Friedel, RH, Gambadoro, A, Gates, H, Hansen, J, Herault, Y, Hicks, GG, Hörlein, A, de Angelis, M Hrabé, Iyer, V, de Jong, PJ, Koscielny, G, Kühn, R, Liu, P, Lloyd, KCK, Lopez, RG, Marschall, S, Martínez, S, McKerlie, C, Meehan, T, von Melchner, H, Moore, M, Murray, SA, Nagy, A, Nutter, LMJ, Pavlovic, G, Pombero, A, Prosser, H, Ramirez-Solis, R, Ringwald, M, Rosen, B, Rosenthal, N, Rossant, J, Noppinger, P Ruiz, Ryder, E, Skarnes, WC, Schick, J, Schnütgen, F, Schofield, P, Seisenberger, C, Selloum, M, Smedley, D, Simpson, EM, Stewart, AF, Teboul, L, Valentini, GP Tocchini, Valenzuela, D, West, AP, and Wurst, W
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Pharmacology and Pharmaceutical Sciences ,Biomedical and Clinical Sciences ,Biotechnology ,Genetics ,Stem Cell Research - Embryonic - Non-Human ,Stem Cell Research ,Human Genome ,2.6 Resources and infrastructure (aetiology) ,Generic health relevance ,Neurosciences ,Neurology & Neurosurgery ,Pharmacology and pharmaceutical sciences - Abstract
The International Knockout Mouse Consortium (IKMC) developed high throughput gene trapping and gene targeting pipelines that produced mostly conditional mutations of more than 18,500 genes in C57BL/6N mouse embryonic stem (ES) cells which have been archived and are freely available to the research community as a frozen resource. From this unprecedented resource more than 6,000 mutant mouse strains have been produced by the IKMC and mostly the International Mouse Phenotyping Consortium (IMPC). In addition, a cre-driver resource was established including 250 inducible cre-driver mouse strains in a C57BL/6 background. Complementing the cre-driver resource, a collection of comprising 27 cre-driver rAAVs has also been produced. The resources can be easily accessed at the IKMC/IMPC web portal (www.mousephenotype.org). The IKMC/IMPC resource is a standardized reference library of mouse models with defined genetic backgrounds that enables the analysis of gene-disease associations in mice of different genetic makeup and should therefore have a major impact on biomedical research.
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- 2016
3. Cleft palate and minor metabolic disturbances in a mouse global Arl15 gene knockout.
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Bai, Y., Bentley, L., Ma, C., Naveenan, N., Cleak, J., Wu, Y., Simon, M.M., Westerberg, H., Cañas, R.C., Horner, N., Pandey, R., Paphiti, K., Schulze, U., Mianné, J., Hough, T., Teboul, L., Baaij, J.H.F. de, Cox, R.D., Bai, Y., Bentley, L., Ma, C., Naveenan, N., Cleak, J., Wu, Y., Simon, M.M., Westerberg, H., Cañas, R.C., Horner, N., Pandey, R., Paphiti, K., Schulze, U., Mianné, J., Hough, T., Teboul, L., Baaij, J.H.F. de, and Cox, R.D.
- Abstract
01 november 2023, Contains fulltext : 296932.pdf (Publisher’s version ) (Open Access), ARL15, a small GTPase protein, was linked to metabolic traits in association studies. We aimed to test the Arl15 gene as a functional candidate for metabolic traits in the mouse. CRISPR/Cas9 germline knockout (KO) of Arl15 showed that homozygotes were postnatal lethal and exhibited a complete cleft palate (CP). Also, decreased cell migration was observed from Arl15 KO mouse embryonic fibroblasts (MEFs). Metabolic phenotyping of heterozygotes showed that females had reduced fat mass on a chow diet from 14 weeks of age. Mild body composition phenotypes were also observed in heterozygous mice on a high-fat diet (HFD)/low-fat diet (LFD). Females on a HFD showed reduced body weight, gonadal fat depot weight and brown adipose tissue (BAT) weight. In contrast, in the LFD group, females showed increased bone mineral density (BMD), while males showed a trend toward reduced BMD. Clinical biochemistry analysis of plasma on HFD showed transient lower adiponectin at 20 weeks of age in females. Urinary and plasma Mg(2+) concentrations were not significantly different. Our phenotyping data showed that Arl15 is essential for craniofacial development. Adult metabolic phenotyping revealed potential roles in brown adipose tissue and bone development.
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- 2023
4. Genome wide conditional mouse knockout resources
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Kaloff, C., Anastassiadis, K., Ayadi, A., Baldock, R., Beig, J., Birling, M.-C., Bradley, A., Brown, S.D.M., Bürger, A., Bushell, W., Chiani, F., Collins, F.S., Doe, B., Eppig, J.T., Finnell, R.H., Fletcher, C., Flicek, P., Fray, M., Friedel, R.H., Gambadoro, A., Gates, H., Hansen, J., Herault, Y., Hicks, G.G., Hörlein, A., Hrabé de Angelis, M., Iyer, V., de Jong, P.J., Koscielny, G., Kühn, R., Liu, P., Lloyd, K.C.K, Lopez, R.G., Marschall, S., Martínez, S., McKerlie, C., Meehan, T., von Melchner, H., Moore, M., Murray, S.A., Nagy, A., Nutter, L.M.J., Pavlovic, G., Pombero, A., Prosser, H., Ramirez-Solis, R., Ringwald, M., Rosen, B., Rosenthal, N., Rossant, J., Ruiz Noppinger, P., Ryder, E., Skarnes, W.C., Schick, J., Schnütgen, F., Schofield, P., Seisenberger, C., Selloum, M., Smedley, D., Simpson, E.M., Stewart, A.F., Teboul, L., Tocchini Valentini, G.P., Valenzuela, D., West, A.P., and Wurst, W.
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- 2016
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5. Broad-spectrum XX and XY gonadal dysgenesis in patients with a homozygous L193S variant in PPP2R3C
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Cicek D, Warr N, Yesil G, H Kocak Eker, Bas F, Poyrazoglu S, Darendeliler F, Direk G, Hatipoglu N, Eltan M, Z Yavas Abali, B Gurpinar Tosun, Kaygusuz SB, T Seven Menevse, Helvacioglu D, Turan S, Bereket A, Reeves R, Simon M, Mackenzie M, Teboul L, Greenfield A, and Guran T
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General Economics, Econometrics and Finance - Published
- 2022
6. Importing genetically altered animals: ensuring quality
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Birling, M.-C., primary, Fray, M. D., additional, Kasparek, P., additional, Kopkanova, J., additional, Massimi, M., additional, Matteoni, R., additional, Montoliu, L., additional, Nutter, L. M. J., additional, Raspa, M., additional, Rozman, J., additional, Ryder, E. J., additional, Scavizzi, F., additional, Voikar, V., additional, Wells, S., additional, Pavlovic, G., additional, and Teboul, L., additional
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- 2021
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7. Le non-sujet de l�antis�mitisme � gauche
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Brenni, Camilla, primary, Krickeberg, Memphis, additional, Nicolas-Teboul, L�a, additional, and Zoubir, Zacharias, additional
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- 2019
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8. Analysis of mammalian gene function through broad-based phenotypic screens across a consortium of mouse clinics
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Hrabe de Angelis, M. (M), Nicholson, G. (G), Selloum, M. (Mohammed), White, J. (J) K. (K), Morgan, H. (H), Ramirez-Solis, R. (R), Sorg, T. (Tania), Wells, S. (S), Fuchs, H. (H), Fray, M. (M), Adams, D. (D) J. (J), Adams, N. (N) C. (C), Adler, T. (T), Aguilar-Pimentel, A. (A), Ali-Hadji, D. (Dalila), Amann, G. (Grégory), Andre, P. (Philippe), Atkins, S. (S), Auburtin, A. (Aurélie), Ayadi, A. (Abdelkader), Becker, J. (Julien), Becker, L. (L), Bedu, E. (Elodie), Bekeredjian, R. (R), Birling, M. (Marie-Christine), Blake, A. (A), Bottomley, J. (J), Bowl, M. (M) R. (R), Brault, V. (Véronique), Busch, D. (D) H. (H), Bussell, J. (J) N. (N), Calzada-Wack, J. (J), Cater, H. (H), Champy, M. (Marie-France), Charles, P. (Philippe), Chevalier, C. (Claire), Chiani, F. (F), Codner, G. (G) F. (F), Combe, R. (R), Cox, R. (R), Dalloneau, E. (E), Dierich, A. (A), Di Fenza, A. (A), Doe, B. (B), Duchon, A. (Arnaud), Eickelberg, O. (O), Esapa, C. (C) T. (T), Fertak, L. (L) E. (E), Feigel, T. (T), Emelyanova, I. (I), Estabel, J. (J), Favor, J. (J), Flenniken, A. (A), Gambadoro, A. (A), Garrett, L. (L), Gates, H. (H), Gerdin, A. (A) K. (K), Gkoutos, G. (G), Greenaway, S. (S), Glasl, L. (L), Goetz, P. (P), Da Cruz, I. (I) G. (G), Gotz, A. (A), Graw, J. (J), Guimond, A. (Alain), Hans, W. (W), Hicks, G. (G), Holter, S. (S) M. (M), Hofler, H. (H), Hancock, J. (J) M. (M), Hoehndorf, R. (R), Hough, T. (T), Houghton, R. (R), Hurt, A. (A), Ivandic, B. (B), Jacobs, H. (Hugues), Jacquot, S. (Sylvie), Jones, N. (N), Karp, N. (N) A. (A), Katus, H. (H) A. (A), Kitchen, S. (S), Klein-Rodewald, T. (T), Klingenspor, M. (M), Klopstock, T. (T), Lalanne, V. (Valérie), Leblanc, S. (Sophie), Lengger, C. (C), le Marchand, E. (Elise), Ludwig, T. (T), Lux, A. (Aline), McKerlie, C. (C), Maier, H. (H), Mandel, J. (Jean-Louis), Marschall, S. (S), Mark, M. (Manuel), Melvin, D. (D) G. (G), Meziane, H. (Hamid), Micklich, K. (K), Mittelhauser, C. (C), Monassier, L. (Laurent), Moulaert, D. (David), Muller, S. (Stéphanie), Naton, B. (B), Neff, F. (F), Nolan, P. (P) M. (M), Nutter, L. (L) M. (M), Ollert, M. (M), Pavlovic, G. (Guillaume), Pellegata, N. (N) S. (S), Peter, E. (E), Petit-Demouliere, B. (Benoît), Pickard, A. (A), Podrini, C. (C), Potter, P. (P), Pouilly, L. (Laurent), Puk, O. (O), Richardson, D. (D), Rousseau, S. (Stéphane), Quintanilla-Fend, L. (L), Quwailid, M. (M) M. (M), Racz, I. (I), Rathkolb, B. (B), Riet, F. (Fabrice), Rossant, J. (J), Roux, M. (Michel), Rozman, J. (J), Ryder, E. (E), Salisbury, J. (J), Santos, L. (L), Schable, K. (K) H. (H), Schiller, E. (E), Schrewe, A. (A), Schulz, H. (H), Steinkamp, R. (R), Simon, M. (M), Stewart, M. (M), Stoger, C. (C), Stoger, T. (T), Sun, M. (M), Sunter, D. (D), Teboul, L. (L), Tilly, I. (I), Tocchini-Valentini, G. (G) P. (P), Tost, M. (M), Treise, I. (I), Vasseur, L. (Laurent), Velot, E. (E), Vogt-Weisenhorn, D. (D), Wagner, C. (Christel), Walling, A. (A), Wattenhofer-Donze, M. (Marie), Weber, B. (Bruno), Wendling, O. (Olivia), Westerberg, H. (H), Willershauser, M. (M), Wolf, E. (E), Wolter, A. (A), Wood, J. (J), Wurst, W. (W), Yildirim, A. (A) O. (O), Zeh, R. (R), Zimmer, A. (A), Zimprich, A. (A), Consortium, E. (Eumodic), Holmes, C. (C), Steel, K. (K) P. (P), Herault, Y. (Yann), Gailus-Durner, V. (V), Mallon, A. (A) M. (M), and Brown, S. (S) D. (D)
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Genetics ,Male ,Mice, Knockout ,Heterozygote ,Mutant ,Homozygote ,Aucun ,Molecular Sequence Annotation ,Biology ,Phenotype ,Mice, Inbred C57BL ,Pleiotropy ,ddc:570 ,Mutation ,Animals ,Humans ,Human genome ,Female ,Allele ,Gene ,Gene knockout ,Genetic Association Studies - Abstract
The function of the majority of genes in the mouse and human genomes remains unknown. The mouse embryonic stem cell knockout resource provides a basis for the characterization of relationships between genes and phenotypes. The EUMODIC consortium developed and validated robust methodologies for the broad-based phenotyping of knockouts through a pipeline comprising 20 disease-oriented platforms. We developed new statistical methods for pipeline design and data analysis aimed at detecting reproducible phenotypes with high power. We acquired phenotype data from 449 mutant alleles, representing 320 unique genes, of which half had no previous functional annotation. We captured data from over 27,000 mice, finding that 83% of the mutant lines are phenodeviant, with 65% demonstrating pleiotropy. Surprisingly, we found significant differences in phenotype annotation according to zygosity. New phenotypes were uncovered for many genes with previously unknown function, providing a powerful basis for hypothesis generation and further investigation in diverse systems. Comment in : Genetic differential calculus. [Nat Genet. 2015] Comment in : Scaling up phenotyping studies. [Nat Biotechnol. 2015]
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- 2015
9. High-resolution Tomographic Imaging Using Coherent Hard x-rays From Compact Laser Driven Accelerators
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Symes, D. R., primary, Najmudin, Z., additional, Cole, J. M., additional, Wood, J. C., additional, Lopes, N.C., additional, Poder, K., additional, Abel, P., additional, Abel, R. L., additional, Alatabi, S., additional, Kneip, S., additional, Mecseki, K., additional, Winkler, M., additional, Foster, P. S., additional, Norris, D. P., additional, Teboul, L., additional, Johnson, S., additional, Szoke-Kovacs, Z., additional, Sandholzer, M., additional, Botchway, S., additional, Gratton, S., additional, Hill, M. A., additional, De Lazzari, M., additional, and Thomson, J., additional
- Published
- 2016
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10. Deficiency of the zinc finger protein ZFP106 causes motor and sensory neurodegeneration.
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Sportbiologie, Joyce PI, Fratta P, Landman AS, Mcgoldrick P, Wackerhage H, Groves M, Busam BS, Galino J, Corrochano S, Beskina OA, Esapa C, Ryder E, Carter S, Stewart M, Codner G, Hilton H, Teboul L, Tucker J, Lionikas A, Estabel J, Ramirez-Solis R, White JK, Brandner S, Plagnol V, Bennet DL, Abramov AY, Greensmith L, Fisher EM, Acevedo-Arozena A, Sportbiologie, and Joyce PI, Fratta P, Landman AS, Mcgoldrick P, Wackerhage H, Groves M, Busam BS, Galino J, Corrochano S, Beskina OA, Esapa C, Ryder E, Carter S, Stewart M, Codner G, Hilton H, Teboul L, Tucker J, Lionikas A, Estabel J, Ramirez-Solis R, White JK, Brandner S, Plagnol V, Bennet DL, Abramov AY, Greensmith L, Fisher EM, Acevedo-Arozena A
- Abstract
Zinc finger motifs are distributed amongst many eukaryotic protein families, directing nucleic acid-protein and protein-protein interactions. Zinc finger protein 106 (ZFP106) has previously been associated with roles in immune response, muscle differentiation, testes development and DNA damage, although little is known about its specific function. To further investigate the function of ZFP106, we performed an in-depth characterization of Zfp106 deficient mice (Zfp106(-/-)), and we report a novel role for ZFP106 in motor and sensory neuronal maintenance and survival. Zfp106(-/-) mice develop severe motor abnormalities, major deficits in muscle strength and histopathological changes in muscle. Intriguingly, despite being highly expressed throughout the central nervous system, Zfp106(-/-) mice undergo selective motor and sensory neuronal and axonal degeneration specific to the spinal cord and peripheral nervous system. Neurodegeneration does not occur during development of Zfp106(-/-) mice, suggesting that ZFP106 is likely required for the maintenance of mature peripheral motor and sensory neurons. Analysis of embryonic Zfp106(-/-) motor neurons revealed deficits in mitochondrial function, with an inhibition of Complex I within the mitochondrial electron transport chain. Our results highlight a vital role for ZFP106 in sensory and motor neuron maintenance and reveal a novel player in mitochondrial dysfunction and neurodegeneration.
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- 2015
11. Deep short-read sequencing of chromosome 17 from the mouse strains A/J and CAST/Ei identifies significant germline variation and candidate genes that regulate liver triglyceride levels
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Sudbery, I., Stalker, J., Simpson, J.T., Keane, T., Rust, A.G., Hurles, M.E., Walter, K., Lynch, D., Teboul, L., Brown, S.D., Li, H., Ning, Z., Nadeau, J.H., Croniger, C.M., Durbin, R., and Adams, D.J.
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animal diseases ,food and beverages - Abstract
Genome sequences are essential tools for comparative and mutational analyses. Here we present the short read sequence of mouse chromosome 17 from the Mus musculus domesticus derived strain A/J, and the Mus musculus castaneus derived strain CAST/Ei. We describe approaches for the accurate identification of nucleotide and structural variation in the genomes of vertebrate experimental organisms, and show how these techniques can be applied to help prioritize candidate genes within quantitative trait loci.
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- 2009
12. The mammalian gene function resource: the international knockout mouse consortium
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Bradley, A, Anastassiadis, K, Ayadi, Azzeddine, Battey, JF, Bell, C, Birling, MC, Bottomley, J, Brown, SD, Burger, A, Bult, CJ, Bushell, W, Collins, FS, Desaintes, C, Doe, B, Economides, A, Eppig, JT, Finnell, RH, D Fletcher, C, Fray, M, Frendewey, D, Friedel, RH, Grosveld, Frank, Hansen, J, Herault, Y, Hicks, G, Horlein, A, Houghton, R, de Angelis, MH, Huylebroeck, Danny, Iyer, V, de Jong, PJ, Kadin, JA, Kaloff, C, Kennedy, K, Koutsourakis, M (Manousos), Lloyd, KCK, Marschall, S, Mason, J, McKerlie, C, McLeod, MP, von Melchner, H, Moore, M, Mujica, AO, Nagy, A, Nefedov, M, Nutter, LM, Pavlovic, G, Peterson, JL, Pollock, J, Ramirez-Solis, R, Rancourt, DE, Raspa, M, Remacle, JE, Ringwald, M, Rosen, B, Rosenthal, N, Rossant, J, Noppinger, PR, Ryder, E, Schick, JZ, Schnutgen, F, Schofield, P, Seisenberger, C, Selloum, M, Simpson, EM, Skarnes, WC, Smedley, D, Stanford, WL, Stewart, AF, Stone, K, Swan, K, Tadepally, H, Teboul, L, Tocchini-Valentini, GP, Valenzuela, D, West, AP, Yamamura, K, Yoshinaga, Y, Wurst, W, Bradley, A, Anastassiadis, K, Ayadi, Azzeddine, Battey, JF, Bell, C, Birling, MC, Bottomley, J, Brown, SD, Burger, A, Bult, CJ, Bushell, W, Collins, FS, Desaintes, C, Doe, B, Economides, A, Eppig, JT, Finnell, RH, D Fletcher, C, Fray, M, Frendewey, D, Friedel, RH, Grosveld, Frank, Hansen, J, Herault, Y, Hicks, G, Horlein, A, Houghton, R, de Angelis, MH, Huylebroeck, Danny, Iyer, V, de Jong, PJ, Kadin, JA, Kaloff, C, Kennedy, K, Koutsourakis, M (Manousos), Lloyd, KCK, Marschall, S, Mason, J, McKerlie, C, McLeod, MP, von Melchner, H, Moore, M, Mujica, AO, Nagy, A, Nefedov, M, Nutter, LM, Pavlovic, G, Peterson, JL, Pollock, J, Ramirez-Solis, R, Rancourt, DE, Raspa, M, Remacle, JE, Ringwald, M, Rosen, B, Rosenthal, N, Rossant, J, Noppinger, PR, Ryder, E, Schick, JZ, Schnutgen, F, Schofield, P, Seisenberger, C, Selloum, M, Simpson, EM, Skarnes, WC, Smedley, D, Stanford, WL, Stewart, AF, Stone, K, Swan, K, Tadepally, H, Teboul, L, Tocchini-Valentini, GP, Valenzuela, D, West, AP, Yamamura, K, Yoshinaga, Y, and Wurst, W
- Abstract
In 2007, the International Knockout Mouse Consortium (IKMC) made the ambitious promise to generate mutations in virtually every protein-coding gene of the mouse genome in a concerted worldwide action. Now, 5 years later, the IKMC members have developed high-throughput gene trapping and, in particular, gene-targeting pipelines and generated more than 17,400 mutant murine embryonic stem (ES) cell clones and more than 1,700 mutant mouse strains, most of them conditional. A common IKMC web portal (www.knockoutmouse.org) has been established, allowing easy access to this unparalleled biological resource. The IKMC materials considerably enhance functional gene annotation of the mammalian genome and will have a major impact on future biomedical research.
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- 2012
13. Difficultés de prise en charge des ulcères veineux : regards croisés médecins généralistes – infirmiers libéraux – patients
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Bensoussan Teboul, L., primary, Renaudin, J.-M., additional, and Bureau, J.-M., additional
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- 2012
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14. AspectTAZ : a new approach based on aspect oriented programming for object oriented industrial messaging services design.
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Teboul, L., Pawlak, R., Seinturier, L., Gressier-Soudan, E., and Becquet, E.
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- 2002
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15. Trans-differentiation of myoblasts to adipoblasts: triggering effects of fatty acids and thiazolidinediones
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Grimaldi, P.A., primary, Teboul, L., additional, Inadera, H., additional, Gaillard, D., additional, and Amri, E.Z., additional
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- 1997
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16. Thiazolidinediones and fatty acids convert myogenic cells into adipose-like cells.
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Teboul, L, Gaillard, D, Staccini, L, Inadera, H, Amri, E Z, and Grimaldi, P A
- Abstract
Fatty acids and thiazolidinediones act as potent activators of the adipose differentiation program in established preadipose cell lines. In this report, the effects of these agents on the differentiation pathway of myoblasts have been investigated. Exposure of C2C12N myoblasts (a subclone of the C2C12 cell line) to thiazolidinediones or fatty acids prevents the expression of myogenin, alpha-actin, and creatine kinase, thus abolishing the formation of multinucleated myotubes. These treatments lead in parallel to the expression of a typical adipose differentiation program including acquisition of adipocyte morphology and activation of adipose-related genes. A similar transition toward the adipose differentiation pathway also occurs in mouse muscle satellite cells maintained in primary culture. Thiazolidinediones exert their adipogenic effects only in non-terminally differentiated myoblasts; myotubes are insensitive to the compounds. Continuous exposure to inducers after growth arrest is not required to maintain the adipose phenotype, but proliferation of adipose-like C2C12N cells leads to a complete reversion toward undifferentiated cells able to undergo either myogenic or adipogenic differentiation depending on the composition of culture medium. These results indicate that adipogenic inducers, such as thiazolidinediones or fatty acids, specifically convert the differentiation pathway of myoblasts into that of adipoblasts.
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- 1995
17. Evidence for a common mechanism of action for fatty acids and thiazolidinedione antidiabetic agents on gene expression in preadipose cells.
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Ibrahimi, A, Teboul, L, Gaillard, D, Amri, E Z, Ailhaud, G, Young, P, Cawthorne, M A, and Grimaldi, P A
- Abstract
In diabetic rodents, thiazolidinediones are able to improve insulin sensitivity of target tissues and to reverse, at least partially, the diabetic state. The effects of these drugs on phenotypic expression in various tissues, including adipose tissue, have been reported. We report here that a new thiazolidinedione compound, BRL 49653, exerts, in preadipose cells, potent effects on the expression of genes encoding proteins involved in fatty acid metabolism. These effects of BRL 49653 in Ob 1771 preadipose cells are similar, in terms of kinetics, reversibility, specificity of genes affected, and requirement for protein synthesis, to those already described for natural or nonmetabolizable fatty acids. Moreover, when used at submaximally effective concentrations, BRL49653 and 2-bromopalmitate act in an additive manner to induce gene expression in preadipose cells, but this additivity of effects is lost when one of the compounds is used at a maximally effective concentration. These observations, suggesting similar mechanisms of action for thiazolidinediones and fatty acids, are strongly supported by the demonstration that (i) both molecules activate, in a heterogolous trans-activation assay, the same nuclear receptor of the steroid/thyroid hormone nuclear receptor superfamily and (ii) transfection of 3T3-C2 fibroblasts with an expression vector for this nuclear receptor confers thiazolidinedione inducibility of adipocyte lipid-binding protein gene expression.
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- 1994
18. Evidence for a common mechanism of action for fatty acids and thiazolidinedione antidiabetic agents on gene expression in preadipose cells
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Ibrahimi A, Teboul L, Gaillard D, Ez, Amri, Gérard Ailhaud, Young P, Ma, Cawthorne, and Pa, Grimaldi
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Fatty Acids ,Palmitates ,Receptors, Cytoplasmic and Nuclear ,Nerve Tissue Proteins ,3T3 Cells ,Fatty Acid-Binding Proteins ,Neoplasm Proteins ,Rats ,Rosiglitazone ,Kinetics ,Mice ,Thiazoles ,Adipose Tissue ,Gene Expression Regulation ,Diabetes Mellitus ,Animals ,Hypoglycemic Agents ,Thiazolidinediones ,RNA, Messenger ,Rats, Wistar ,Carrier Proteins ,Fatty Acid-Binding Protein 7 ,Cells, Cultured - Abstract
In diabetic rodents, thiazolidinediones are able to improve insulin sensitivity of target tissues and to reverse, at least partially, the diabetic state. The effects of these drugs on phenotypic expression in various tissues, including adipose tissue, have been reported. We report here that a new thiazolidinedione compound, BRL 49653, exerts, in preadipose cells, potent effects on the expression of genes encoding proteins involved in fatty acid metabolism. These effects of BRL 49653 in Ob 1771 preadipose cells are similar, in terms of kinetics, reversibility, specificity of genes affected, and requirement for protein synthesis, to those already described for natural or nonmetabolizable fatty acids. Moreover, when used at submaximally effective concentrations, BRL49653 and 2-bromopalmitate act in an additive manner to induce gene expression in preadipose cells, but this additivity of effects is lost when one of the compounds is used at a maximally effective concentration. These observations, suggesting similar mechanisms of action for thiazolidinediones and fatty acids, are strongly supported by the demonstration that (i) both molecules activate, in a heterogolous trans-activation assay, the same nuclear receptor of the steroid/thyroid hormone nuclear receptor superfamily and (ii) transfection of 3T3-C2 fibroblasts with an expression vector for this nuclear receptor confers thiazolidinedione inducibility of adipocyte lipid-binding protein gene expression.
19. Correction of the auditory phenotype in C57BL/6N mice via CRISPR/Cas9-mediated homology directed repair
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Mianné J, Lauren Chessum, Kumar S, Aguilar C, Codner G, Hutchison M, Parker A, Am, Mallon, Wells S, Mm, Simon, Teboul L, Sd, Brown, and Bowl MR
20. p27Kip1 independently promotes neuronal differentiation and migration in the cerebral cortex,p27Kip1 independently promotes neuronal differentiation and migration in the cerebral cortex
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Nguyen, L., Besson, A., Julian Ik-Tsen Heng, Schuurmans, C., Teboul, L., Parras, C., Philpott, A., Roberts, J. M., and Guillemot, F.
21. AspectTAZ : a new approach based on aspect oriented programming for object oriented industrial messaging services design
- Author
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Teboul, L., primary, Pawlak, R., additional, Seinturier, L., additional, Gressier-Soudan, E., additional, and Becquet, E., additional
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22. Impact of essential genes on the success of genome editing experiments generating 3313 new genetically engineered mouse lines.
- Author
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Elrick H, Peterson KA, Willis BJ, Lanza DG, Acar EF, Ryder EJ, Teboul L, Kasparek P, Birling MC, Adams DJ, Bradley A, Braun RE, Brown SD, Caulder A, Codner GF, DeMayo FJ, Dickinson ME, Doe B, Duddy G, Gertsenstein M, Goodwin LO, Hérault Y, Lintott LG, Lloyd KCK, Lorenzo I, Mackenzie M, Mallon AM, McKerlie C, Parkinson H, Ramirez-Solis R, Seavitt JR, Sedlacek R, Skarnes WC, Smedley D, Wells S, White JK, Wood JA, Murray SA, Heaney JD, and Nutter LMJ
- Subjects
- Animals, Mice, CRISPR-Cas Systems, Alleles, Mice, Inbred C57BL, Male, Female, Genetic Engineering methods, Phenotype, Genes, Essential, Gene Editing methods, Mice, Knockout
- Abstract
The International Mouse Phenotyping Consortium (IMPC) systematically produces and phenotypes mouse lines with presumptive null mutations to provide insight into gene function. The IMPC now uses the programmable RNA-guided nuclease Cas9 for its increased capacity and flexibility to efficiently generate null alleles in the C57BL/6N strain. In addition to being a valuable novel and accessible research resource, the production of 3313 knockout mouse lines using comparable protocols provides a rich dataset to analyze experimental and biological variables affecting in vivo gene engineering with Cas9. Mouse line production has two critical steps - generation of founders with the desired allele and germline transmission (GLT) of that allele from founders to offspring. A systematic evaluation of the variables impacting success rates identified gene essentiality as the primary factor influencing successful production of null alleles. Collectively, our findings provide best practice recommendations for using Cas9 to generate alleles in mouse essential genes, many of which are orthologs of genes linked to human disease., (© 2024. The Author(s).)
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- 2024
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23. Improving laboratory animal genetic reporting: LAG-R guidelines.
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Teboul L, Amos-Landgraf J, Benavides FJ, Birling MC, Brown SDM, Bryda E, Bunton-Stasyshyn R, Chin HJ, Crispo M, Delerue F, Dobbie M, Franklin CL, Fuchtbauer EM, Gao X, Golzio C, Haffner R, Hérault Y, Hrabe de Angelis M, Lloyd KCK, Magnuson TR, Montoliu L, Murray SA, Nam KH, Nutter LMJ, Pailhoux E, Pardo Manuel de Villena F, Peterson K, Reinholdt L, Sedlacek R, Seong JK, Shiroishi T, Smith C, Takeo T, Tinsley L, Vilotte JL, Warming S, Wells S, Whitelaw CB, Yoshiki A, and Pavlovic G
- Subjects
- Animals, Reproducibility of Results, Research Design, Animal Experimentation standards, Biomedical Research standards, Animals, Laboratory genetics, Guidelines as Topic
- Abstract
The biomedical research community addresses reproducibility challenges in animal studies through standardized nomenclature, improved experimental design, transparent reporting, data sharing, and centralized repositories. The ARRIVE guidelines outline documentation standards for laboratory animals in experiments, but genetic information is often incomplete. To remedy this, we propose the Laboratory Animal Genetic Reporting (LAG-R) framework. LAG-R aims to document animals' genetic makeup in scientific publications, providing essential details for replication and appropriate model use. While verifying complete genetic compositions may be impractical, better reporting and validation efforts enhance reliability of research. LAG-R standardization will bolster reproducibility, peer review, and overall scientific rigor., (© 2024. The Author(s).)
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- 2024
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24. Identification of cellular retinoic acid binding protein 2 (CRABP2) as downstream target of nuclear factor I/X (NFIX): implications for skeletal dysplasia syndromes.
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Kooblall KG, Stevenson M, Heilig R, Stewart M, Wright B, Lockstone H, Buck D, Fischer R, Wells S, Lines KE, Teboul L, Hennekam RC, and Thakker RV
- Abstract
Nuclear factor I/X ( NFIX ) mutations are associated with 2 skeletal dysplasias, Marshall-Smith (MSS) and Malan (MAL) syndromes. NFIX encodes a transcription factor that regulates expression of genes, including Bobby sox ( BBX ) and glial fibrillary acidic protein ( GFAP ) in neural progenitor cells and astrocytes, respectively. To elucidate the role of NFIX mutations in MSS, we studied their effects in fibroblast cell lines obtained from 5 MSS unrelated patients and 3 unaffected individuals. The 5 MSS NFIX frameshift mutations in exons 6-8 comprised 3 deletions (c.819-732_1079-948del, c.819-471_1079-687del, c.819-592_1079-808del), an insertion (c.1037_1038insT), and a duplication (c.1090dupG). Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analyses using MSS and unrelated control fibroblasts and in vitro expression studies in monkey kidney fibroblast (COS-7) cells showed that frameshift mutations in NFIX exons 6-8 generated mutant transcripts that were not cleared by nonsense-mediated-decay mechanisms and encoded truncated NFIX proteins. Moreover, BBX or GFAP expression was unaffected in the majority of MSS fibroblasts. To identify novel NFIX downstream target genes, RNA sequencing and proteomics analyses were performed on mouse embryonic fibroblast (MEF) cells derived from control Nfix
+/+ , Nfix+/Del2 , Nfix+/Del24 , NfixDel24/Del24 , Nfix+/Del140 , and NfixDel140/Del140 mice, compared with NfixDel2/Del2 mice which had developmental, skeletal, and neural abnormalities. This identified 191 transcripts and 815 proteins misregulated in NfixDel2/Del2 MEFs with ≥2-fold-change ( P <0 .05). Validation studies using qRT-PCR and western blot analyses confirmed that 2 genes, cellular retinoic acid binding protein 2 ( Crabp2 ) and vascular cell adhesion molecule 1 ( Vcam1 ), were misregulated at the RNA and protein levels in NfixDel2/Del2 MEFs, and that CRABP2 and VCAM1 expressions were altered in 60%-100% of MSS fibroblast cells. Furthermore, in vitro luciferase reporter assays confirmed that NFIX directly regulates CRABP2 promoter activity. Thus, these altered genes and pathways may represent possible targets for drugs as potential treatments and therapies for MSS., Competing Interests: The authors declare no competing interests., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society for Bone and Mineral Research.)- Published
- 2024
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25. Long-read sequencing for fast and robust identification of correct genome-edited alleles: PCR-based and Cas9 capture methods.
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McCabe CV, Price PD, Codner GF, Allan AJ, Caulder A, Christou S, Loeffler J, Mackenzie M, Malzer E, Mianné J, Nowicki KJ, O'Neill EJ, Pike FJ, Hutchison M, Petit-Demoulière B, Stewart ME, Gates H, Wells S, Sanderson ND, and Teboul L
- Subjects
- Animals, Alleles, Recombinational DNA Repair, Polymerase Chain Reaction, CRISPR-Cas Systems genetics, Gene Editing methods
- Abstract
Background: Recent developments in CRISPR/Cas9 genome-editing tools have facilitated the introduction of precise alleles, including genetic intervals spanning several kilobases, directly into the embryo. However, the introduction of donor templates, via homology directed repair, can be erroneous or incomplete and these techniques often produce mosaic founder animals. Thus, newly generated alleles must be verified at the sequence level across the targeted locus. Screening for the presence of the desired mutant allele using traditional sequencing methods can be challenging due to the size of the interval to be sequenced, together with the mosaic nature of founders., Methodology/principal Findings: In order to help disentangle the genetic complexity of these animals, we tested the application of Oxford Nanopore Technologies long-read sequencing at the targeted locus and found that the achievable depth of sequencing is sufficient to offset the sequencing error rate associated with the technology used to validate targeted regions of interest. We have assembled an analysis workflow that facilitates interrogating the entire length of a targeted segment in a single read, to confirm that the intended mutant sequence is present in both heterozygous animals and mosaic founders. We used this workflow to compare the output of PCR-based and Cas9 capture-based targeted sequencing for validation of edited alleles., Conclusion: Targeted long-read sequencing supports in-depth characterisation of all experimental models that aim to produce knock-in or conditional alleles, including those that contain a mix of genome-edited alleles. PCR- or Cas9 capture-based modalities bring different advantages to the analysis., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: LT was the recipient of a conference travel award from ONT., (Copyright: © 2024 McCabe et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2024
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26. How much do we know about the function of mammalian genes?
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Teboul L, Hérault Y, Wells S, and Pavlovic G
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- Animals, Mammals genetics
- Published
- 2023
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27. Shiga toxin targets the podocyte causing hemolytic uremic syndrome through endothelial complement activation.
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Bowen EE, Hurcombe JA, Barrington F, Keir LS, Farmer LK, Wherlock MD, Ortiz-Sandoval CG, Bruno V, Bohorquez-Hernandez A, Diatlov D, Rostam-Shirazi N, Wells S, Stewart M, Teboul L, Lay AC, Butler MJ, Pope RJP, Larkai EMS, Morgan BP, Moppett J, Satchell SC, Welsh GI, Walker PD, Licht C, Saleem MA, and Coward RJM
- Subjects
- Child, Humans, Mice, Animals, Shiga Toxin genetics, Shiga Toxin metabolism, Shiga Toxin therapeutic use, Vascular Endothelial Growth Factor A metabolism, Vascular Endothelial Growth Factor A therapeutic use, Complement Activation, Podocytes metabolism, Podocytes pathology, Escherichia coli Infections complications, Escherichia coli Infections drug therapy, Escherichia coli Infections metabolism, Hemolytic-Uremic Syndrome drug therapy, Hemolytic-Uremic Syndrome metabolism, Hemolytic-Uremic Syndrome pathology, Shiga-Toxigenic Escherichia coli metabolism, Kidney Diseases pathology
- Abstract
Background: Shiga toxin (Stx)-producing Escherichia coli hemolytic uremic syndrome (STEC-HUS) is the leading cause of acute kidney injury in children, with an associated mortality of up to 5%. The mechanisms underlying STEC-HUS and why the glomerular microvasculature is so susceptible to injury following systemic Stx infection are unclear., Methods: Transgenic mice were engineered to express the Stx receptor (Gb3) exclusively in their kidney podocytes (Pod-Gb3) and challenged with systemic Stx. Human glomerular cell models and kidney biopsies from patients with STEC-HUS were also studied., Findings: Stx-challenged Pod-Gb3 mice developed STEC-HUS. This was mediated by a reduction in podocyte vascular endothelial growth factor A (VEGF-A), which led to loss of glomerular endothelial cell (GEnC) glycocalyx, a reduction in GEnC inhibitory complement factor H binding, and local activation of the complement pathway. Early therapeutic inhibition of the terminal complement pathway with a C5 inhibitor rescued this podocyte-driven, Stx-induced HUS phenotype., Conclusions: This study potentially explains why systemic Stx exposure targets the glomerulus and supports the early use of terminal complement pathway inhibition in this devastating disease., Funding: This work was supported by the UK Medical Research Council (MRC) (grant nos. G0901987 and MR/K010492/1) and Kidney Research UK (grant nos. TF_007_20151127, RP42/2012, and SP/FSGS1/2013). The Mary Lyon Center is part of the MRC Harwell Institute and is funded by the MRC (A410)., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2023
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28. Cleft palate and minor metabolic disturbances in a mouse global Arl15 gene knockout.
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Bai Y, Bentley L, Ma C, Naveenan N, Cleak J, Wu Y, Simon MM, Westerberg H, Cañas RC, Horner N, Pandey R, Paphiti K, Schulze U, Mianné J, Hough T, Teboul L, de Baaij JHF, and Cox RD
- Subjects
- Male, Female, Mice, Animals, Gene Knockout Techniques, Fibroblasts metabolism, Diet, High-Fat, Adipose Tissue, Brown metabolism, Adiponectin metabolism, Mice, Inbred C57BL, Mice, Knockout, Cleft Palate genetics, Cleft Palate metabolism
- Abstract
ARL15, a small GTPase protein, was linked to metabolic traits in association studies. We aimed to test the Arl15 gene as a functional candidate for metabolic traits in the mouse. CRISPR/Cas9 germline knockout (KO) of Arl15 showed that homozygotes were postnatal lethal and exhibited a complete cleft palate (CP). Also, decreased cell migration was observed from Arl15 KO mouse embryonic fibroblasts (MEFs). Metabolic phenotyping of heterozygotes showed that females had reduced fat mass on a chow diet from 14 weeks of age. Mild body composition phenotypes were also observed in heterozygous mice on a high-fat diet (HFD)/low-fat diet (LFD). Females on a HFD showed reduced body weight, gonadal fat depot weight and brown adipose tissue (BAT) weight. In contrast, in the LFD group, females showed increased bone mineral density (BMD), while males showed a trend toward reduced BMD. Clinical biochemistry analysis of plasma on HFD showed transient lower adiponectin at 20 weeks of age in females. Urinary and plasma Mg
2+ concentrations were not significantly different. Our phenotyping data showed that Arl15 is essential for craniofacial development. Adult metabolic phenotyping revealed potential roles in brown adipose tissue and bone development., (© 2023 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)- Published
- 2023
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29. A Mouse Model with a Frameshift Mutation in the Nuclear Factor I/X ( NFIX ) Gene Has Phenotypic Features of Marshall-Smith Syndrome.
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Kooblall KG, Stevenson M, Stewart M, Harris L, Zalucki O, Dewhurst H, Butterfield N, Leng H, Hough TA, Ma D, Siow B, Potter P, Cox RD, Brown SDM, Horwood N, Wright B, Lockstone H, Buck D, Vincent TL, Hannan FM, Bassett JHD, Williams GR, Lines KE, Piper M, Wells S, Teboul L, Hennekam RC, and Thakker RV
- Abstract
The nuclear factor I/X ( NFIX ) gene encodes a ubiquitously expressed transcription factor whose mutations lead to two allelic disorders characterized by developmental, skeletal, and neural abnormalities, namely, Malan syndrome (MAL) and Marshall-Smith syndrome (MSS). NFIX mutations associated with MAL mainly cluster in exon 2 and are cleared by nonsense-mediated decay (NMD) leading to NFIX haploinsufficiency, whereas NFIX mutations associated with MSS are clustered in exons 6-10 and escape NMD and result in the production of dominant-negative mutant NFIX proteins. Thus, different NFIX mutations have distinct consequences on NFIX expression. To elucidate the in vivo effects of MSS-associated NFIX exon 7 mutations, we used CRISPR-Cas9 to generate mouse models with exon 7 deletions that comprised: a frameshift deletion of two nucleotides ( Nfix Del2); in-frame deletion of 24 nucleotides ( Nfix Del24); and deletion of 140 nucleotides ( Nfix Del140). Nfix
+/Del2 , Nfix+/Del24 , Nfix+/Del140 , NfixDel24/Del24 , and NfixDel140/Del140 mice were viable, normal, and fertile, with no skeletal abnormalities, but NfixDel2/Del2 mice had significantly reduced viability ( p < 0.002) and died at 2-3 weeks of age. Nfix Del2 was not cleared by NMD, and NfixDel2/Del2 mice, when compared to Nfix+/+ and Nfix+/Del2 mice, had: growth retardation; short stature with kyphosis; reduced skull length; marked porosity of the vertebrae with decreased vertebral and femoral bone mineral content; and reduced caudal vertebrae height and femur length. Plasma biochemistry analysis revealed NfixDel2/Del2 mice to have increased total alkaline phosphatase activity but decreased C-terminal telopeptide and procollagen-type-1-N-terminal propeptide concentrations compared to Nfix+/+ and Nfix+/Del2 mice. NfixDel2/Del2 mice were also found to have enlarged cerebral cortices and ventricular areas but smaller dentate gyrus compared to Nfix+/+ mice. Thus, NfixDel2/Del2 mice provide a model for studying the in vivo effects of NFIX mutants that escape NMD and result in developmental abnormalities of the skeletal and neural tissues that are associated with MSS. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research., (© 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.)- Published
- 2023
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30. Excess of guide RNA reduces knockin efficiency and drastically increases on-target large deletions.
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Chenouard V, Leray I, Tesson L, Remy S, Allan A, Archer D, Caulder A, Fortun A, Bernardeau K, Cherifi Y, Teboul L, David L, and Anegon I
- Abstract
CRISPR-Cas9 cleavage efficacy and accuracy are the main challenges gene editing faces, and they are particularly affected by the optimal formation of the ribonucleoprotein (RNP) complex. We used nano differential scanning fluorimetry, a label and immobilization-free assay, to demonstrate that an equimolar ratio of Cas9 and guide RNA (gRNA) is optimal for RNP complex formation. We almost achieved 50% of green fluorescent protein (GFP) to blue fluorescent protein (BFP) conversion using a biallelic homozygous GFP human induced pluripotent stem cell line, when 0.4 μM of Cas9, equimolar Cas9/gRNA ratio and 2 μM of single-stranded oligonucleotide, were used and showed that increasing Cas9/gRNA ratio did not further improve KI efficiency. Additionally, excess gRNA decreased point mutation KI efficiency in rat embryos and drastically increased the occurrence of on-target large deletions. These findings highlight the importance of CRISPR/Cas9 stoichiometric optimization to ensure efficient and accurate KI generation, which will be applicable to other in vitro as well as in vivo models., Competing Interests: YC and VC were genOway employees. The other authors declare no conflict of interest., (© 2023 The Authors.)
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- 2023
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31. TMEM161B regulates cerebral cortical gyration, Sonic Hedgehog signaling, and ciliary structure in the developing central nervous system.
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Akula SK, Marciano JH, Lim Y, Exposito-Alonso D, Hylton NK, Hwang GH, Neil JE, Dominado N, Bunton-Stasyshyn RK, Song JHT, Talukdar M, Schmid A, Teboul L, Mo A, Shin T, Finander B, Beck SG, Yeh RC, Otani A, Qian X, DeGennaro EM, Alkuraya FS, Maddirevula S, Cascino GD, Giannini C, Burrage LC, Rosenfield JA, Ketkar S, Clark GD, Bacino C, Lewis RA, Segal RA, Bazan JF, Smith KA, Golden JA, Cho G, and Walsh CA
- Subjects
- Animals, Female, Humans, Mice, Pregnancy, Central Nervous System metabolism, Cilia genetics, Cilia metabolism, Mice, Knockout, Signal Transduction, Ferrets, Hedgehog Proteins genetics, Hedgehog Proteins metabolism
- Abstract
Sonic hedgehog signaling regulates processes of embryonic development across multiple tissues, yet factors regulating context-specific Shh signaling remain poorly understood. Exome sequencing of families with polymicrogyria (disordered cortical folding) revealed multiple individuals with biallelic deleterious variants in TMEM161B , which encodes a multi-pass transmembrane protein of unknown function. Tmem161b null mice demonstrated holoprosencephaly, craniofacial midline defects, eye defects, and spinal cord patterning changes consistent with impaired Shh signaling, but were without limb defects, suggesting a CNS-specific role of Tmem161b. Tmem161b depletion impaired the response to Smoothened activation in vitro and disrupted cortical histogenesis in vivo in both mouse and ferret models, including leading to abnormal gyration in the ferret model. Tmem161b localizes non-exclusively to the primary cilium, and scanning electron microscopy revealed shortened, dysmorphic, and ballooned ventricular zone cilia in the Tmem161b null mouse, suggesting that the Shh-related phenotypes may reflect ciliary dysfunction. Our data identify TMEM161B as a regulator of cerebral cortical gyration, as involved in primary ciliary structure, as a regulator of Shh signaling, and further implicate Shh signaling in human gyral development.
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- 2023
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32. Genotyping Genome-Edited Founders and Subsequent Generation.
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Mackenzie M, Fower A, Allan AJ, Codner GF, Bunton-Stasyshyn RK, and Teboul L
- Subjects
- Animals, Genotype, Mutation, Genome, CRISPR-Cas Systems, Gene Editing
- Abstract
Targeted nucleases allow the production of many types of genetic mutations directly in the early embryo. However, the outcome of their activity is a repair event of unpredictable nature, and the founder animals that are produced are generally of a mosaic nature. Here, we present the molecular assays and genotyping strategies that will support the screening of the first generation for potential founders and the validation of positive animals in the subsequent generation, according to the type of mutation generated., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
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- 2023
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33. Mendelian gene identification through mouse embryo viability screening.
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Cacheiro P, Westerberg CH, Mager J, Dickinson ME, Nutter LMJ, Muñoz-Fuentes V, Hsu CW, Van den Veyver IB, Flenniken AM, McKerlie C, Murray SA, Teboul L, Heaney JD, Lloyd KCK, Lanoue L, Braun RE, White JK, Creighton AK, Laurin V, Guo R, Qu D, Wells S, Cleak J, Bunton-Stasyshyn R, Stewart M, Harrisson J, Mason J, Haseli Mashhadi H, Parkinson H, Mallon AM, and Smedley D
- Subjects
- Animals, Female, Homozygote, Humans, Mice, Mice, Knockout, Phenotype, Pregnancy, Embryo, Mammalian, Genes, Lethal
- Abstract
Background: The diagnostic rate of Mendelian disorders in sequencing studies continues to increase, along with the pace of novel disease gene discovery. However, variant interpretation in novel genes not currently associated with disease is particularly challenging and strategies combining gene functional evidence with approaches that evaluate the phenotypic similarities between patients and model organisms have proven successful. A full spectrum of intolerance to loss-of-function variation has been previously described, providing evidence that gene essentiality should not be considered as a simple and fixed binary property., Methods: Here we further dissected this spectrum by assessing the embryonic stage at which homozygous loss-of-function results in lethality in mice from the International Mouse Phenotyping Consortium, classifying the set of lethal genes into one of three windows of lethality: early, mid, or late gestation lethal. We studied the correlation between these windows of lethality and various gene features including expression across development, paralogy and constraint metrics together with human disease phenotypes. We explored a gene similarity approach for novel gene discovery and investigated unsolved cases from the 100,000 Genomes Project., Results: We found that genes in the early gestation lethal category have distinct characteristics and are enriched for genes linked with recessive forms of inherited metabolic disease. We identified several genes sharing multiple features with known biallelic forms of inborn errors of the metabolism and found signs of enrichment of biallelic predicted pathogenic variants among early gestation lethal genes in patients recruited under this disease category. We highlight two novel gene candidates with phenotypic overlap between the patients and the mouse knockouts., Conclusions: Information on the developmental period at which embryonic lethality occurs in the knockout mouse may be used for novel disease gene discovery that helps to prioritise variants in unsolved rare disease cases., (© 2022. The Author(s).)
- Published
- 2022
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34. Erratum: Generation and analysis of innovative genomically humanized knockin SOD1 , TARDBP (TDP-43), and FUS mouse models.
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Devoy A, Price G, De Giorgio F, Bunton-Stasyshyn R, Thompson D, Gasco S, Allan A, Codner GF, Nair RR, Tibbit C, McLeod R, Ali Z, Noda J, Marrero-Gagliardi A, Brito-Armas JM, Williams C, Öztürk MM, Simon M, O'Neill E, Bryce-Smith S, Harrison J, Atkins G, Corrochano S, Stewart M, Gilthorpe JD, Teboul L, Acevedo-Arozena A, Fisher EMC, and Cunningham TJ
- Abstract
[This corrects the article DOI: 10.1016/j.isci.2021.103463.]., (© 2022 The Author(s).)
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- 2022
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35. Introduction to Mammalian Genome Special Issue: Mammalian Genetic Resources.
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Brown SDM, Nadeau JH, Wells S, Mallon AM, Teboul L, Tuggle CK, and Schook LB
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- Animals, Genome, Mammals genetics
- Published
- 2022
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36. Extensive identification of genes involved in congenital and structural heart disorders and cardiomyopathy.
- Author
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Spielmann N, Miller G, Oprea TI, Hsu CW, Fobo G, Frishman G, Montrone C, Haseli Mashhadi H, Mason J, Munoz Fuentes V, Leuchtenberger S, Ruepp A, Wagner M, Westphal DS, Wolf C, Görlach A, Sanz-Moreno A, Cho YL, Teperino R, Brandmaier S, Sharma S, Galter IR, Östereicher MA, Zapf L, Mayer-Kuckuk P, Rozman J, Teboul L, Bunton-Stasyshyn RKA, Cater H, Stewart M, Christou S, Westerberg H, Willett AM, Wotton JM, Roper WB, Christiansen AE, Ward CS, Heaney JD, Reynolds CL, Prochazka J, Bower L, Clary D, Selloum M, Bou About G, Wendling O, Jacobs H, Leblanc S, Meziane H, Sorg T, Audain E, Gilly A, Rayner NW, Hitz MP, Zeggini E, Wolf E, Sedlacek R, Murray SA, Svenson KL, Braun RE, White JK, Kelsey L, Gao X, Shiroishi T, Xu Y, Seong JK, Mammano F, Tocchini-Valentini GP, Beaudet AL, Meehan TF, Parkinson H, Smedley D, Mallon AM, Wells SE, Grallert H, Wurst W, Marschall S, Fuchs H, Brown SDM, Flenniken AM, Nutter LMJ, McKerlie C, Herault Y, Lloyd KCK, Dickinson ME, Gailus-Durner V, and Hrabe de Angelis M
- Abstract
Clinical presentation of congenital heart disease is heterogeneous, making identification of the disease-causing genes and their genetic pathways and mechanisms of action challenging. By using in vivo electrocardiography, transthoracic echocardiography and microcomputed tomography imaging to screen 3,894 single-gene-null mouse lines for structural and functional cardiac abnormalities, here we identify 705 lines with cardiac arrhythmia, myocardial hypertrophy and/or ventricular dilation. Among these 705 genes, 486 have not been previously associated with cardiac dysfunction in humans, and some of them represent variants of unknown relevance (VUR). Mice with mutations in Casz1, Dnajc18, Pde4dip, Rnf38 or Tmem161b genes show developmental cardiac structural abnormalities, with their human orthologs being categorized as VUR. Using UK Biobank data, we validate the importance of the DNAJC18 gene for cardiac homeostasis by showing that its loss of function is associated with altered left ventricular systolic function. Our results identify hundreds of previously unappreciated genes with potential function in congenital heart disease and suggest causal function of five VUR in congenital heart disease., (© 2022. The Author(s).)
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- 2022
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37. Screening and validation of genome-edited animals.
- Author
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Bunton-Stasyshyn RK, Codner GF, and Teboul L
- Subjects
- Alleles, Animals, Embryo, Mammalian, Reproducibility of Results, Gene Editing methods, Gene Editing standards, Genetic Testing
- Abstract
The emergence of an array of genome-editing tools in recent years has facilitated the introduction of genetic modifications directly into the embryo, increasing the ease, efficiency and catalogue of alleles accessible to researchers across a range of species. Bypassing the requirement for a selection cassette and resulting in a broad range of outcomes besides the desired allele, genome editing has altered the allele validation process both temporally and technically. Whereas traditional gene targeting relies upon selection and allows allele validation at the embryonic stem cell modification stage, screening for the presence of the intended allele now occurs in the (frequently mosaic) founder animals. Final confirmation of the edited allele can only take place at the subsequent G1 generation and the validation strategy must differentiate the desired allele from a range of unintended outcomes. Here we present some of the challenges posed by gene editing, strategies for validation and considerations for animal colony management.
- Published
- 2022
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38. Neuroplastin genetically interacts with Cadherin 23 and the encoded isoform Np55 is sufficient for cochlear hair cell function and hearing.
- Author
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Newton S, Kong F, Carlton AJ, Aguilar C, Parker A, Codner GF, Teboul L, Wells S, Brown SDM, Marcotti W, and Bowl MR
- Subjects
- Animals, Loss of Function Mutation, Mice, Mice, Knockout, Plasma Membrane Calcium-Transporting ATPases metabolism, Cadherins genetics, Hair Cells, Auditory, Inner physiology, Hearing genetics, Membrane Glycoproteins genetics, Protein Isoforms genetics
- Abstract
Mammalian hearing involves the mechanoelectrical transduction (MET) of sound-induced fluid waves in the cochlea. Essential to this process are the specialised sensory cochlear cells, the inner (IHCs) and outer hair cells (OHCs). While genetic hearing loss is highly heterogeneous, understanding the requirement of each gene will lead to a better understanding of the molecular basis of hearing and also to therapeutic opportunities for deafness. The Neuroplastin (Nptn) gene, which encodes two protein isoforms Np55 and Np65, is required for hearing, and homozygous loss-of-function mutations that affect both isoforms lead to profound deafness in mice. Here we have utilised several distinct mouse models to elaborate upon the spatial, temporal, and functional requirement of Nptn for hearing. While we demonstrate that both Np55 and Np65 are present in cochlear cells, characterisation of a Np65-specific mouse knockout shows normal hearing thresholds indicating that Np65 is functionally redundant for hearing. In contrast, we find that Nptn-knockout mice have significantly reduced maximal MET currents and MET channel open probabilities in mature OHCs, with both OHCs and IHCs also failing to develop fully mature basolateral currents. Furthermore, comparing the hearing thresholds and IHC synapse structure of Nptn-knockout mice with those of mice that lack Nptn only in IHCs and OHCs shows that the majority of the auditory deficit is explained by hair cell dysfunction, with abnormal afferent synapses contributing only a small proportion of the hearing loss. Finally, we show that continued expression of Neuroplastin in OHCs of adult mice is required for membrane localisation of Plasma Membrane Ca2+ ATPase 2 (PMCA2), which is essential for hearing function. Moreover, Nptn haploinsufficiency phenocopies Atp2b2 (encodes PMCA2) mutations, with heterozygous Nptn-knockout mice exhibiting hearing loss through genetic interaction with the Cdh23ahl allele. Together, our findings provide further insight to the functional requirement of Neuroplastin for mammalian hearing., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2022
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39. Broad-spectrum XX and XY gonadal dysgenesis in patients with a homozygous L193S variant in PPP2R3C.
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Cicek D, Warr N, Yesil G, Kocak Eker H, Bas F, Poyrazoglu S, Darendeliler F, Direk G, Hatipoglu N, Eltan M, Yavas Abali Z, Gurpinar Tosun B, Kaygusuz SB, Seven Menevse T, Helvacioglu D, Turan S, Bereket A, Reeves R, Simon M, Mackenzie M, Teboul L, Greenfield A, and Guran T
- Subjects
- Amino Acid Substitution, Animals, Child, Consanguinity, Embryo, Mammalian, Female, Gonadal Dysgenesis, 46,XX pathology, Gonadal Dysgenesis, 46,XY pathology, Homozygote, Humans, Leucine genetics, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Mutation, Missense, Pedigree, Pregnancy, Serine genetics, Gonadal Dysgenesis, 46,XX genetics, Gonadal Dysgenesis, 46,XY genetics, Protein Phosphatase 2 genetics
- Abstract
Context: Homozygous and heterozygous variants in PPP2R3C are associated with syndromic 46,XY complete gonadal dysgenesis (Myo-Ectodermo-Gonadal Dysgenesis (MEGD) syndrome), and impaired spermatogenesis, respectively. This study expands the role of PPP2R3C in the aetiology of gonadal dysgenesis (GD)., Method: We sequenced the PPP2R3C gene in four new patients from three unrelated families. The clinical, laboratory, and molecular characteristics were investigated. We have also determined the requirement for Ppp2r3c in mice (C57BL6/N) using CRISPR/Cas9 genome editing., Results: A homozygous c.578T>C (p.L193S) PPP2R3C variant was identified in one 46,XX girl with primary gonadal insufficiency, two girls with 46,XY complete GD, and one undervirilised boy with 46,XY partial GD. The patients with complete GD had low gonadal and adrenal androgens, low anti-Müllerian hormone, and high follicle-stimulating hormone and luteinizing hormone concentrations. All patients manifested characteristic features of MEGD syndrome. Heterozygous Ppp2r3c knockout mice appeared overtly normal and fertile. Inspection of homozygous embryos at 14.5, 9.5, and 8.5 days post coitum(dpc) revealed evidence of dead embryos. We conclude that loss of function of Ppp2r3c is not compatible with viability in mice and results in embryonic death from 7.5 dpc or earlier., Conclusion: Our data indicate the essential roles for PPP2R3C in mouse and human development. Germline homozygous variants in human PPP2R3C are associated with distinctive syndromic GD of varying severity in both 46,XY and 46,XX individuals.
- Published
- 2021
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40. Generation and analysis of innovative genomically humanized knockin SOD1 , TARDBP (TDP-43), and FUS mouse models.
- Author
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Devoy A, Price G, De Giorgio F, Bunton-Stasyshyn R, Thompson D, Gasco S, Allan A, Codner GF, Nair RR, Tibbit C, McLeod R, Ali Z, Noda J, Marrero-Gagliardi A, Brito-Armas JM, Williams C, Öztürk MM, Simon M, O'Neill E, Bryce-Smith S, Harrison J, Atkins G, Corrochano S, Stewart M, Gilthorpe JD, Teboul L, Acevedo-Arozena A, Fisher EMC, and Cunningham TJ
- Abstract
Amyotrophic lateral sclerosis/frontotemporal dementia (ALS/FTD) is a fatal neurodegenerative disorder, and continued innovation is needed for improved understanding and for developing therapeutics. We have created next-generation genomically humanized knockin mouse models, by replacing the mouse genomic region of Sod1 , Tardbp (TDP-43), and Fus , with their human orthologs, preserving human protein biochemistry and splicing with exons and introns intact. We establish a new standard of large knockin allele quality control, demonstrating the utility of indirect capture for enrichment of a genomic region of interest followed by Oxford Nanopore sequencing. Extensive analysis shows that homozygous humanized animals only express human protein at endogenous levels. Characterization of humanized FUS animals showed that they are phenotypically normal throughout their lifespan. These humanized strains are vital for preclinical assessment of interventions and serve as templates for the addition of coding or non-coding human ALS/FTD mutations to dissect disease pathomechanisms, in a physiological context., Competing Interests: The authors declare no competing interests., (© 2021 The Authors.)
- Published
- 2021
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41. Linking the FTO obesity rs1421085 variant circuitry to cellular, metabolic, and organismal phenotypes in vivo.
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Laber S, Forcisi S, Bentley L, Petzold J, Moritz F, Smirnov KS, Al Sadat L, Williamson I, Strobel S, Agnew T, Sengupta S, Nicol T, Grallert H, Heier M, Honecker J, Mianne J, Teboul L, Dumbell R, Long H, Simon M, Lindgren C, Bickmore WA, Hauner H, Schmitt-Kopplin P, Claussnitzer M, and Cox RD
- Subjects
- Adipocytes metabolism, Animals, Diet, High-Fat adverse effects, Male, Mice, Phenotype, Polymorphism, Single Nucleotide, Alpha-Ketoglutarate-Dependent Dioxygenase FTO genetics, Alpha-Ketoglutarate-Dependent Dioxygenase FTO metabolism, Obesity genetics, Obesity metabolism
- Abstract
Variants in FTO have the strongest association with obesity; however, it is still unclear how those noncoding variants mechanistically affect whole-body physiology. We engineered a deletion of the rs1421085 conserved cis-regulatory module (CRM) in mice and confirmed in vivo that the CRM modulates Irx3 and Irx5 gene expression and mitochondrial function in adipocytes. The CRM affects molecular and cellular phenotypes in an adipose depot-dependent manner and affects organismal phenotypes that are relevant for obesity, including decreased high-fat diet-induced weight gain, decreased whole-body fat mass, and decreased skin fat thickness. Last, we connected the CRM to a genetically determined effect on steroid patterns in males that was dependent on nutritional challenge and conserved across mice and humans. Together, our data establish cross-species conservation of the rs1421085 regulatory circuitry at the molecular, cellular, metabolic, and organismal level, revealing previously unknown contextual dependence of the variant's action., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).)
- Published
- 2021
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42. Methods of genome engineering and model validation.
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Teboul L
- Subjects
- Genetic Engineering, Genome, Bacterial, Synthetic Biology, Genome genetics
- Published
- 2021
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43. Universal Southern blot protocol with cold or radioactive probes for the validation of alleles obtained by homologous recombination.
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Codner GF, Erbs V, Loeffler J, Chessum L, Caulder A, Jullien N, Wells S, Birling MC, and Teboul L
- Subjects
- Alleles, Animals, Blotting, Southern, Gene Targeting, Genetic Vectors, Homologous Recombination, Mice, Radioactivity
- Abstract
The widespread availability of recombineered vectors and gene targeted embryonic stem cells from large-scale repositories facilitates the generation of mouse models for functional genetic studies. Southern blotting validates the structure of these targeted alleles produced by homologous recombination, as well as indicating any additional integrations of the vector into the genome. Traditionally this technique employs radioactively-labelled probes; however, there are many laboratories that are restricted in their use of radioactivity. Here, we present a widely applicable protocol for Southern blot analysis using cold probes and alternative procedures employing radioactive probes. Furthermore, the probes are designed to recognise standardised regions of gene-targeting cassettes and so represent universally applicable reagents for assessing allelic integrity., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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44. Ap2s1 mutation causes hypercalcaemia in mice and impairs interaction between calcium-sensing receptor and adaptor protein-2.
- Author
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Hannan FM, Stevenson M, Bayliss AL, Stokes VJ, Stewart M, Kooblall KG, Gorvin CM, Codner G, Teboul L, Wells S, and Thakker RV
- Subjects
- Animals, Bone Density genetics, CRISPR-Cas Systems genetics, Calcium metabolism, Cinacalcet pharmacology, Disease Models, Animal, Gene Editing, Humans, Hypercalcemia drug therapy, Hypercalcemia metabolism, Hypercalcemia pathology, Mice, Mutation genetics, Phenotype, Adaptor Protein Complex 2 genetics, Adaptor Protein Complex sigma Subunits genetics, Fibroblast Growth Factor-23 genetics, Hypercalcemia genetics, Receptors, Calcium-Sensing genetics
- Abstract
Adaptor protein 2 (AP2), a heterotetrameric complex comprising AP2α, AP2β2, AP2μ2 and AP2σ2 subunits, is ubiquitously expressed and involved in endocytosis and trafficking of membrane proteins, such as the calcium-sensing receptor (CaSR), a G-protein coupled receptor that signals via Gα11. Mutations of CaSR, Gα11 and AP2σ2, encoded by AP2S1, cause familial hypocalciuric hypercalcaemia types 1-3 (FHH1-3), respectively. FHH3 patients have heterozygous AP2S1 missense Arg15 mutations (p.Arg15Cys, p.Arg15His or p.Arg15Leu) with hypercalcaemia, which may be marked and symptomatic, and occasional hypophosphataemia and osteomalacia. To further characterize the phenotypic spectrum and calcitropic pathophysiology of FHH3, we used CRISPR/Cas9 genome editing to generate mice harboring the AP2S1 p.Arg15Leu mutation, which causes the most severe FHH3 phenotype. Heterozygous (Ap2s1+/L15) mice were viable, and had marked hypercalcaemia, hypermagnesaemia, hypophosphataemia, and increases in alkaline phosphatase activity and fibroblast growth factor-23. Plasma 1,25-dihydroxyvitamin D was normal, and no alterations in bone mineral density or bone turnover were noted. Homozygous (Ap2s1L15/L15) mice invariably died perinatally. Co-immunoprecipitation studies showed that the AP2S1 p.Arg15Leu mutation impaired protein-protein interactions between AP2σ2 and the other AP2 subunits, and also with the CaSR. Cinacalcet, a CaSR positive allosteric modulator, decreased plasma calcium and parathyroid hormone concentrations in Ap2s1+/L15 mice, but had no effect on the diminished AP2σ2-CaSR interaction in vitro. Thus, our studies have established a mouse model that is representative for FHH3 in humans, and demonstrated that the AP2S1 p.Arg15Leu mutation causes a predominantly calcitropic phenotype, which can be ameliorated by treatment with cinacalcet., (© The Author(s) 2021. Published by Oxford University Press.)
- Published
- 2021
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45. A novel knockout mouse for the small EDRK-rich factor 2 (Serf2) showing developmental and other deficits.
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Cleverley K, Lee WC, Mumford P, Collins T, Rickman M, Cunningham TJ, Cleak J, Mianne J, Szoke-Kovacs Z, Stewart M, Teboul L, Maduro C, Wells S, Wiseman FK, and Fisher EMC
- Subjects
- Age Factors, Alleles, Alternative Splicing, Animals, Cell Line, Disease Models, Animal, Embryonic Stem Cells metabolism, Female, Gene Expression Regulation, Genetic Background, Genetic Loci, Genotype, Male, Mice, Mice, Knockout, Organ Specificity, X-Ray Microtomography, Developmental Disabilities diagnosis, Developmental Disabilities genetics, Genetic Association Studies methods, Genetic Predisposition to Disease, Intracellular Signaling Peptides and Proteins genetics, Phenotype
- Abstract
The small EDRK-rich factor 2 (SERF2) is a highly conserved protein that modifies amyloid fibre assembly in vitro and promotes protein misfolding. However, the role of SERF2 in regulating age-related proteotoxicity remains largely unexplored due to a lack of in vivo models. Here, we report the generation of Serf2 knockout mice using an ES cell targeting approach, with Serf2 knockout alleles being bred onto different defined genetic backgrounds. We highlight phenotyping data from heterozygous Serf2
+/- mice, including unexpected male-specific phenotypes in startle response and pre-pulse inhibition. We report embryonic lethality in Serf2-/- null animals when bred onto a C57BL/6 N background. However, homozygous null animals were viable on a mixed genetic background and, remarkably, developed without obvious abnormalities. The Serf2 knockout mice provide a powerful tool to further investigate the role of SERF2 protein in previously unexplored pathophysiological pathways in the context of a whole organism.- Published
- 2021
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- View/download PDF
46. A resource of targeted mutant mouse lines for 5,061 genes.
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Birling MC, Yoshiki A, Adams DJ, Ayabe S, Beaudet AL, Bottomley J, Bradley A, Brown SDM, Bürger A, Bushell W, Chiani F, Chin HG, Christou S, Codner GF, DeMayo FJ, Dickinson ME, Doe B, Donahue LR, Fray MD, Gambadoro A, Gao X, Gertsenstein M, Gomez-Segura A, Goodwin LO, Heaney JD, Hérault Y, de Angelis MH, Jiang ST, Justice MJ, Kasparek P, King RE, Kühn R, Lee H, Lee YJ, Liu Z, Lloyd KCK, Lorenzo I, Mallon AM, McKerlie C, Meehan TF, Fuentes VM, Newman S, Nutter LMJ, Oh GT, Pavlovic G, Ramirez-Solis R, Rosen B, Ryder EJ, Santos LA, Schick J, Seavitt JR, Sedlacek R, Seisenberger C, Seong JK, Skarnes WC, Sorg T, Steel KP, Tamura M, Tocchini-Valentini GP, Wang CL, Wardle-Jones H, Wattenhofer-Donzé M, Wells S, Wiles MV, Willis BJ, Wood JA, Wurst W, Xu Y, Teboul L, and Murray SA
- Subjects
- Animals, Information Dissemination, International Cooperation, Internet, Mice, Mice, Knockout, Mouse Embryonic Stem Cells cytology, Mouse Embryonic Stem Cells metabolism, Mutagenesis, Phenotype, Gene Deletion, Genetic Association Studies, Genome, Genotype
- Published
- 2021
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47. LAMA: automated image analysis for the developmental phenotyping of mouse embryos.
- Author
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Horner NR, Venkataraman S, Armit C, Casero R, Brown JM, Wong MD, van Eede MC, Henkelman RM, Johnson S, Teboul L, Wells S, Brown SD, Westerberg H, and Mallon AM
- Subjects
- Animals, Female, Imaging, Three-Dimensional methods, Mice, Mice, Inbred C57BL, Mice, Knockout physiology, Phenotype, Software, Embryo, Mammalian physiology, Image Processing, Computer-Assisted methods
- Abstract
Advanced 3D imaging modalities, such as micro-computed tomography (micro-CT), have been incorporated into the high-throughput embryo pipeline of the International Mouse Phenotyping Consortium (IMPC). This project generates large volumes of raw data that cannot be immediately exploited without significant resources of personnel and expertise. Thus, rapid automated annotation is crucial to ensure that 3D imaging data can be integrated with other multi-dimensional phenotyping data. We present an automated computational mouse embryo phenotyping pipeline that harnesses the large amount of wild-type control data available in the IMPC embryo pipeline in order to address issues of low mutant sample number as well as incomplete penetrance and variable expressivity. We also investigate the effect of developmental substage on automated phenotyping results. Designed primarily for developmental biologists, our software performs image pre-processing, registration, statistical analysis and segmentation of embryo images. We also present a novel anatomical E14.5 embryo atlas average and, using it with LAMA, show that we can uncover known and novel dysmorphology from two IMPC knockout lines., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2021. Published by The Company of Biologists Ltd.)
- Published
- 2021
- Full Text
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48. Loss of O-GlcNAcase catalytic activity leads to defects in mouse embryogenesis.
- Author
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Muha V, Authier F, Szoke-Kovacs Z, Johnson S, Gallagher J, McNeilly A, McCrimmon RJ, Teboul L, and van Aalten DMF
- Subjects
- Animals, Catalytic Domain, Embryonic Development genetics, Female, Histone Acetyltransferases metabolism, Histone Acetyltransferases physiology, Homeostasis, Male, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, N-Acetylglucosaminyltransferases metabolism, Pregnancy, Protein Processing, Post-Translational, X-Ray Microtomography methods, beta-N-Acetylhexosaminidases genetics, beta-N-Acetylhexosaminidases physiology, Embryonic Development physiology, beta-N-Acetylhexosaminidases metabolism
- Abstract
O-GlcNAcylation is an essential post-translational modification that has been implicated in neurodevelopmental and neurodegenerative disorders. O-GlcNAcase (OGA), the sole enzyme catalyzing the removal of O-GlcNAc from proteins, has emerged as a potential drug target. OGA consists of an N-terminal OGA catalytic domain and a C-terminal pseudo histone acetyltransferase (HAT) domain with unknown function. To investigate phenotypes specific to loss of OGA catalytic activity and dissect the role of the HAT domain, we generated a constitutive knock-in mouse line, carrying a mutation of a catalytic aspartic acid to alanine. These mice showed perinatal lethality and abnormal embryonic growth with skewed Mendelian ratios after day E18.5. We observed tissue-specific changes in O-GlcNAc homeostasis regulation to compensate for loss of OGA activity. Using X-ray microcomputed tomography on late gestation embryos, we identified defects in the kidney, brain, liver, and stomach. Taken together, our data suggest that developmental defects during gestation may arise upon prolonged OGA inhibition specifically because of loss of OGA catalytic activity and independent of the function of the HAT domain., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
49. Mouse mutant phenotyping at scale reveals novel genes controlling bone mineral density.
- Author
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Swan AL, Schütt C, Rozman J, Del Mar Muñiz Moreno M, Brandmaier S, Simon M, Leuchtenberger S, Griffiths M, Brommage R, Keskivali-Bond P, Grallert H, Werner T, Teperino R, Becker L, Miller G, Moshiri A, Seavitt JR, Cissell DD, Meehan TF, Acar EF, Lelliott CJ, Flenniken AM, Champy MF, Sorg T, Ayadi A, Braun RE, Cater H, Dickinson ME, Flicek P, Gallegos J, Ghirardello EJ, Heaney JD, Jacquot S, Lally C, Logan JG, Teboul L, Mason J, Spielmann N, McKerlie C, Murray SA, Nutter LMJ, Odfalk KF, Parkinson H, Prochazka J, Reynolds CL, Selloum M, Spoutil F, Svenson KL, Vales TS, Wells SE, White JK, Sedlacek R, Wurst W, Lloyd KCK, Croucher PI, Fuchs H, Williams GR, Bassett JHD, Gailus-Durner V, Herault Y, Mallon AM, Brown SDM, Mayer-Kuckuk P, and Hrabe de Angelis M
- Subjects
- Animals, Female, Gene Ontology, Genetic Pleiotropy, Genome-Wide Association Study, Genotype, Male, Mice, Mice, Transgenic, Mutation, Osteoblasts pathology, Osteoclasts pathology, Osteoporosis metabolism, Phenotype, Promoter Regions, Genetic, Protein Interaction Maps, Sex Characteristics, Transcriptome, Bone Density genetics, Gene Expression Regulation genetics, Osteoblasts metabolism, Osteoclasts metabolism, Osteoporosis genetics
- Abstract
The genetic landscape of diseases associated with changes in bone mineral density (BMD), such as osteoporosis, is only partially understood. Here, we explored data from 3,823 mutant mouse strains for BMD, a measure that is frequently altered in a range of bone pathologies, including osteoporosis. A total of 200 genes were found to significantly affect BMD. This pool of BMD genes comprised 141 genes with previously unknown functions in bone biology and was complementary to pools derived from recent human studies. Nineteen of the 141 genes also caused skeletal abnormalities. Examination of the BMD genes in osteoclasts and osteoblasts underscored BMD pathways, including vesicle transport, in these cells and together with in silico bone turnover studies resulted in the prioritization of candidate genes for further investigation. Overall, the results add novel pathophysiological and molecular insight into bone health and disease., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2020
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50. Anticipating and Identifying Collateral Damage in Genome Editing.
- Author
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Burgio G and Teboul L
- Subjects
- Animals, Humans, Risk Assessment, CRISPR-Cas Systems, Gene Editing standards, Genome
- Abstract
Genome editing has powerful applications in research, healthcare, and agriculture. However, the range of possible molecular events resulting from genome editing has been underestimated and the technology remains unpredictable on, and away from, the target locus. This has considerable impact in providing a safe approach for therapeutic genome editing, agriculture, and other applications. This opinion article discusses how to anticipate and detect those editing events by a combination of assays to capture all possible genomic changes. It also discusses strategies for preventing unwanted effects, critical to appraise the benefit or risk associated with the use of the technology. Anticipating and verifying the result of genome editing are essential for the success for all applications., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
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