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3 results on '"Natalia Ewa Janowicz"'

1. Cas9-mediated gene disruption in tetraploid Giardia intestinalis

Author

Filip Weisz, Martina Vinopalová, Pavla Tůmová, Kari D. Hagen, Pavel Doležal, Markéta Petrů, Vendula Horáčková, Natalia Ewa Janowicz, Lenka Marková, Scott C. Dawson, Alžběta Motyčková, and Luboš Voleman

Subjects
Genetics, Transcription (biology), Cas9, Essential gene, CRISPR, Guide RNA, Biology, Homologous recombination, Gene, Genome
Abstract

CRISPR/Cas9 system is an extremely powerful technique that is extensively used for various genome modifications in different organisms including parasitic protists. Giardia intestinalis, a protist parasite infecting about 280 million people around the world each year, has been eluding the routine use of CRISPR/Cas9 for generating knock-out cell lines due to the presence of four copies of each gene in its two nuclei. Apart from single exception employing rather laborious Cre/loxP system, no full knock-out cell line has been established yet. In this work, we show the ability of in-vitro assembled CRISPR/Cas9 components to successfully edit the genome of G. intestinalis. We further established a cell line stably expressing Cas9 in both G. intestinalis nuclei. Subsequent introduction of a template for homologous recombination containing the transcription units for the resistance marker and gRNA resulted in the removal of all gene copies at once for three independent experimental genes, mem, cwp1 and mlf1. The method was also applicable for the incomplete disruption of an essential gene, as documented by markedly decreased expression of tom40. Finally, testing the efficiency of Cas9-induced recombination revealed that homologous arms as short as 150 bp can be sufficient to establish a full knock-out cell line in G. intestinalis.

Published
2021

2. Genetic tool development in marine protists: Emerging model organisms for experimental cell biology

Author

Deepak Nanjappa, Iñaki Ruiz-Trillo, Christopher J. Howe, Chris Bowler, Mariusz Nowacki, Anastasios D. Tsaousis, Noelia Lander, Christopher L. Dupont, Shinichiro Maruyama, Fulei Luan, Andrew E. Allen, Anna M. G. Novák Vanclová, Pamela A. Silver, Nastasia J. Freyria, Peter von Dassow, Elisabeth Hehenberger, Konomi Fujimura-Kamada, Julius Lukeš, Roberto Docampo, Deborah L. Robertson, Thomas E. Clemente, Sebastián R. Najle, Kathryn J. Coyne, Cristina Aresté, Brittany N. Sprecher, Lu Wang, Eleanna Kazana, Verónica Freire-Benéitez, Vladimír Hampl, Cecilia Balestreri, Isabel C. Nimmo, Mariana Rius, Yoshihisa Hirakawa, Arnab Pain, Jorge Ibañez, Elin Einarsson, Lev Tsypin, Heriberto Cerutti, Adrian C. Barbrook, G. Jason Smith, Alexandra Z. Worden, Jian Guo, Jean Claude Lozano, Virginia P. Edgcomb, Huan Zhang, Andrea Highfield, Isaac Núñez, Fatma Gomaa, Jan Pyrih, Natalia Ewa Janowicz, Sara J. Bender, Ross F. Waller, Tobias von der Haar, François-Yves Bouget, Matus Valach, Amanda Hopes, Luke M. Noble, Paulo A. Garcia, Nicholas A.T. Irwin, Tamara Matute, Jernej Turnšek, Ian Hu, Sandra Pucciarelli, Fernán Federici, Valérie Vergé, R. Ellen R. Nisbet, Miguel Angel Chiurillo, Mark Moosburner, Monika Abedin Sigg, Aaron P. Turkewitz, Albane Ruaud, Angela Piersanti, Sebastian G. Gornik, Peter R. Girguis, Adam C. Jones, Lawrence A. Klobutcher, Claudio H. Slamovits, Elena Casacuberta, Imen Lassadi, Elizabeth C. Cooney, Kodai Fukuda, Nicole King, Manuel Ares, Jonathan Z. Kaye, Lisa Sudek, Patrick Beardslee, Cristina Miceli, Xiaoxue Wen, Estienne C. Swart, Yuu Ishii, Ambar Kachale, Pia A. Elustondo, Esteban R. Haro-Contreras, Patrick J. Keeling, José A. Fernández Robledo, Glen L. Wheeler, Colin Brownlee, Rowena Stern, Joshua S. Rest, Susana A. Breglia, Senjie Lin, Duncan B. Coles, Jackie L. Collier, Drahomíra Faktorová, David S. Booth, April Woods, Binnypreet Kaur, Thomas Mock, Gertraud Burger, Jun Minagawa, Yutaka Hanawa, Zhu-Hong Li, Rachele Cesaroni, Laboratoire d'Océanographie Microbienne (LOMIC), Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Institut national des sciences de l'Univers (INSU - CNRS)-Observatoire océanologique de Banyuls (OOB), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Faktorová, Drahomíra [0000-0001-9623-2233], Nisbet, R. Ellen R. [0000-0003-4487-196X], Allen, Andrew E. [0000-0001-5911-6081], Ares, Manuel, Jr [0000-0002-2552-9168], Bowler, Chris [0000-0003-3835-6187], Burger, Gertraud [0000-0002-8679-8812], Collier, Jackie L. [0000-0001-8774-5715], Cooney, Elizabeth C. [0000-0001-7435-7609], Docampo, Roberto [0000-0003-4229-8784], Dupont, Christopher L. [0000-0002-0896-6542], Edgcomb, Virginia [0000-0001-6805-381X], Freyria, Nastasia J. [0000-0002-4972-9383], Gornik, Sebastian G. [0000-0002-8026-1336], Howe, Christopher J. [0000-0002-6975-8640], Hu, Ian [0000-0002-2287-031X], Ishii, Yuu [0000-0003-1735-9557], Jones, Adam C. [0000-0001-7521-1863], Lin, Senjie [0000-0001-8831-6111], Maruyama, Shinichiro [0000-0002-1128-5916], Minagawa, Jun [0000-0002-3028-3203], Najle, Sebastián R. [0000-0002-0654-9147], Nowacki, Mariusz [0000-0003-4894-2905], Pain, Arnab [0000-0002-1755-2819], Silver, Pamela A. [0000-0002-7856-4071], Jason Smith, G. [0000-0003-1258-4800], Sprecher, Brittany N. [0000-0001-5032-3834], Tsaousis, Anastasios D. [0000-0002-5424-1905], Tsypin, Lev [0000-0002-0642-8468], Turkewitz, Aaron [0000-0003-3531-5806], Turnšek, Jernej [0000-0002-9056-3565], Valach, Matus [0000-0001-8689-0080], von der Haar, Tobias [0000-0002-6031-9254], Mock, Thomas [0000-0001-9604-0362], Worden, Alexandra Z. [0000-0002-9888-9324], Lukeš, Julius [0000-0002-0578-6618], Apollo - University of Cambridge Repository, Nisbet, R Ellen R [0000-0003-4487-196X], Allen, Andrew E [0000-0001-5911-6081], Ares, Manuel [0000-0002-2552-9168], Collier, Jackie L [0000-0001-8774-5715], Cooney, Elizabeth C [0000-0001-7435-7609], Dupont, Christopher L [0000-0002-0896-6542], Freyria, Nastasia J [0000-0002-4972-9383], Gornik, Sebastian G [0000-0002-8026-1336], Howe, Christopher J [0000-0002-6975-8640], Jones, Adam C [0000-0001-7521-1863], Najle, Sebastián R [0000-0002-0654-9147], Silver, Pamela A [0000-0002-7856-4071], Jason Smith, G [0000-0003-1258-4800], Sprecher, Brittany N [0000-0001-5032-3834], Tsaousis, Anastasios D [0000-0002-5424-1905], Worden, Alexandra Z [0000-0002-9888-9324], Gordon and Betty Moore Foundation, Leverhulme Trust, Ministry of Education, Youth and Sports (Czech Republic), and European Commission

Subjects
Resource, Molecular biology, Range (biology), Tree of life (biology), [SDV]Life Sciences [q-bio], Green Fluorescent Proteins, ved/biology.organism_classification_rank.species, Marine Biology, Environment, Diversification (marketing strategy), Biology, Models, Biological, Biochemistry, Genome, 03 medical and health sciences, Transformation, Genetic, 0302 clinical medicine, Species Specificity, Genome editing, 631/1647/767/722, Ecosystem, 14. Life underwater, Model organism, Molecular Biology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Dna delivery, 030306 microbiology, ved/biology, Cellular pathways, Eukaryota, Genetic systems, Biodiversity, DNA, Cell Biology, Genetic models, biology.organism_classification, Cell biology, QR, Transformation (genetics), Eukaryote, 030217 neurology & neurosurgery, 631/337, Biotechnology
Abstract

Diverse microbial ecosystems underpin life in the sea. Among these microbes are many unicellular eukaryotes that span the diversity of the eukaryotic tree of life. However, genetic tractability has been limited to a few species, which do not represent eukaryotic diversity or environmentally relevant taxa. Here, we report on the development of genetic tools in a range of protists primarily from marine environments. We present evidence for foreign DNA delivery and expression in 13 species never before transformed and for advancement of tools for eight other species, as well as potential reasons for why transformation of yet another 17 species tested was not achieved. Our resource in genetic manipulation will provide insights into the ancestral eukaryotic lifeforms, general eukaryote cell biology, protein diversification and the evolution of cellular pathways., This collaborative effort was supported by the Gordon and Betty Moore Foundation EMS Program of the Marine Microbiology Initiative (grant nos. GBMF4972 and 4972.01 to F.-Y.B.; GBMF4970 and 4970.01 to M.A. and A.Z.W.; GBMF3788 to A.Z.W.; GBMF 4968 and 4968.01 to H.C.; GBMF4984 to V.H.; GBMF4974 and 4974.01 to C. Brownlee; GBMF4964 to Y. Hirakawa; GBMF4961 to T. Mock; GBMF4958 to P.S.; GBMF4957 to A.T.; GBMF4960 to G.J.S.; GBMF4979 to K.C.; GBMF4982 and 4982.01 to J.L.C.; GBMF4964 to P.J.K.; GBMF4981 to P.v.D.; GBMF5006 to A.E.A.; GBMF4986 to C.M.; GBMF4962 to J.A.F.R.; GBMF4980 and 4980.01 to S.L.; GBMF 4977 and 4977.01 to R.F.W.; GBMF4962.01 to C.H.S.; GBMF4985 to J.M.; GBMF4976 and 4976.01 to C.H.; GBMF4963 and 4963.01 to V.E.; GBMF5007 to C.L.D.; GBMF4983 and 4983.01 to J.L.; GBMF4975 and 4975.01 to A.D.T.; GBMF4973 and 4973.01 to I.R.-T. and GBMF4965 to N.K.), by The Leverhulme Trust (RPG-2017-364) to T. Mock and A. Hopes, and by ERD funds (16_019/0000759) from the Czech Ministry of Education to J.L.

Published
2020

3. Publisher Correction: Genetic tool development in marine protists: emerging model organisms for experimental cell biology

Author

Claudio H. Slamovits, Gertraud Burger, Patrick J. Keeling, Jian Guo, Fatma Gomaa, Jan Pyrih, Jun Minagawa, Christopher L. Dupont, Verónica Freire-Benéitez, Nastasia J. Freyria, Rachele Cesaroni, Elizabeth C. Cooney, Eleanna Kazana, Aaron P. Turkewitz, Peter R. Girguis, Konomi Fujimura-Kamada, Chris Bowler, Anastasios D. Tsaousis, Pamela A. Silver, Lawrence A. Klobutcher, Tamara Matute, Shinichiro Maruyama, Nicole King, Virginia P. Edgcomb, Roberto Docampo, Isaac Núñez, Deborah L. Robertson, Xiaoxue Wen, Zhu-Hong Li, Huan Zhang, Jorge Ibañez, Kathryn J. Coyne, Jonathan Z. Kaye, Christopher J. Howe, Andrew E. Allen, Ross F. Waller, Brittany N. Sprecher, R. Ellen R. Nisbet, Mark Moosburner, Alexandra Z. Worden, Thomas Mock, Angela Piersanti, Sebastian G. Gornik, Paulo A. Garcia, Amanda Hopes, Isabel C. Nimmo, Mariana Rius, Estienne C. Swart, Nicholas A.T. Irwin, Elisabeth Hehenberger, Valérie Vergé, Andrea Highfield, Sandra Pucciarelli, François-Yves Bouget, Yutaka Hanawa, Matus Valach, April Woods, Elin Einarsson, Anna M. G. Novák Vanclová, Lu Wang, Fernán Federici, Monika Abedin Sigg, Adrian C. Barbrook, Yoshihisa Hirakawa, G. Jason Smith, Adam C. Jones, Deepak Nanjappa, Vladimír Hampl, Binnypreet Kaur, Thomas E. Clemente, Jernej Turnšek, Jean Claude Lozano, Sara J. Bender, Ian Hu, Sebastián R. Najle, Imen Lassadi, Glen L. Wheeler, Cecilia Balestreri, Fulei Luan, Lev Tsypin, Heriberto Cerutti, Miguel Angel Chiurillo, Colin Brownlee, Rowena Stern, Julius Lukeš, Manuel Ares, Drahomíra Faktorová, Kodai Fukuda, David S. Booth, Peter von Dassow, Elena Casacuberta, Patrick Beardslee, Cristina Miceli, Cristina Aresté, Natalia Ewa Janowicz, Joshua S. Rest, Susana A. Breglia, Senjie Lin, Luke M. Noble, Albane Ruaud, Duncan B. Coles, Pia A. Elustondo, Jackie L. Collier, Lisa Sudek, Yuu Ishii, José A. Fernández Robledo, Noelia Lander, Tobias von der Haar, Ambar Kachale, Esteban R. Haro-Contreras, Arnab Pain, Iñaki Ruiz-Trillo, and Mariusz Nowacki

Subjects
biology, Molecular biology, media_common.quotation_subject, Garcia, Green Fluorescent Proteins, Drahomira, Eukaryota, Marine Biology, Cell Biology, Art, Biodiversity, DNA, Environment, biology.organism_classification, Genetic models, Biochemistry, Publisher Correction, Models, Biological, Transformation, Genetic, Species Specificity, Humanities, Ecosystem, Biotechnology, media_common
Abstract

Author(s): Faktorova, Drahomira; Nisbet, R Ellen R; Fernandez Robledo, Jose A; Casacuberta, Elena; Sudek, Lisa; Allen, Andrew E; Ares, Manuel; Areste, Cristina; Balestreri, Cecilia; Barbrook, Adrian C; Beardslee, Patrick; Bender, Sara; Booth, David S; Bouget, Francois-Yves; Bowler, Chris; Breglia, Susana A; Brownlee, Colin; Burger, Gertraud; Cerutti, Heriberto; Cesaroni, Rachele; Chiurillo, Miguel A; Clemente, Thomas; Coles, Duncan B; Collier, Jackie L; Cooney, Elizabeth C; Coyne, Kathryn; Docampo, Roberto; Dupont, Christopher L; Edgcomb, Virginia; Einarsson, Elin; Elustondo, Pia A; Federici, Fernan; Freire-Beneitez, Veronica; Freyria, Nastasia J; Fukuda, Kodai; Garcia, Paulo A; Girguis, Peter R; Gomaa, Fatma; Gornik, Sebastian G; Guo, Jian; Hampl, Vladimir; Hanawa, Yutaka; Haro-Contreras, Esteban R; Hehenberger, Elisabeth; Highfield, Andrea; Hirakawa, Yoshihisa; Hopes, Amanda; Howe, Christopher J; Hu, Ian; Ibanez, Jorge; Irwin, Nicholas AT; Ishii, Yuu; Janowicz, Natalia Ewa; Jones, Adam C; Kachale, Ambar; Fujimura-Kamada, Konomi; Kaur, Binnypreet; Kaye, Jonathan Z; Kazana, Eleanna; Keeling, Patrick J; King, Nicole; Klobutcher, Lawrence A; Lander, Noelia; Lassadi, Imen; Li, Zhuhong; Lin, Senjie; Lozano, Jean-Claude; Luan, Fulei; Maruyama, Shinichiro; Matute, Tamara; Miceli, Cristina; Minagawa, Jun; Moosburner, Mark; Najle, Sebastian R; Nanjappa, Deepak; Nimmo, Isabel C; Noble, Luke; Novak Vanclova, Anna MG; Nowacki, Mariusz; Nunez, Isaac; Pain, Arnab; Piersanti, Angela; Pucciarelli, Sandra; Pyrih, Jan; Rest, Joshua S | Abstract: An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Published
2020

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