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Your search keyword '"CSCA TR1"' showing total 10 results
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10 results on '"CSCA TR1"'

1. Cloning-free CRISPR

Author

Arbab, Mandana, Srinivasan, Sharanya, Hashimoto, Tatsunori, Geijsen, Niels, Sherwood, Richard I, Onderzoek, CSCA TR1, Regenerative Medicine, Stem Cells & Cancer, Massachusetts Institute of Technology. Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Srinivasan, Sharanya, Hashimoto, Tatsunori Benjamin, Onderzoek, CSCA TR1, Regenerative Medicine, Stem Cells & Cancer, and Hubrecht Institute for Developmental Biology and Stem Cell Research

Subjects
Resource, Cells, Transgene, Biology, Research Support, Biochemistry, N.I.H, 03 medical and health sciences, Mice, 0302 clinical medicine, Plasmid, Research Support, N.I.H., Extramural, Genetics, Journal Article, CRISPR, Animals, Humans, Cloning, Molecular, Non-U.S. Gov't, lcsh:QH301-705.5, Gene, Gene knockout, Cells, Cultured, 030304 developmental biology, lcsh:R5-920, 0303 health sciences, Cultured, Cas9, Research Support, Non-U.S. Gov't, Extramural, Molecular, Gene targeting, Cell Biology, HEK293 Cells, lcsh:Biology (General), Gene Targeting, CRISPR-Cas Systems, lcsh:Medicine (General), Homologous recombination, 030217 neurology & neurosurgery, Cloning, Developmental Biology
Abstract

We present self-cloning CRISPR/Cas9 (scCRISPR), a technology that allows for CRISPR/Cas9-mediated genomic mutation and site-specific knockin transgene creation within several hours by circumventing the need to clone a site-specific single-guide RNA (sgRNA) or knockin homology construct for each target locus. We introduce a self-cleaving palindromic sgRNA plasmid and a short double-stranded DNA sequence encoding the desired locus-specific sgRNA into target cells, allowing them to produce a locus-specific sgRNA plasmid through homologous recombination. scCRISPR enables efficient generation of gene knockouts (∼88% mutation rate) at approximately one-sixth the cost of plasmid-based sgRNA construction with only 2 hr of preparation for each targeted site. Additionally, we demonstrate efficient site-specific knockin of GFP transgenes without any plasmid cloning or genome-integrated selection cassette in mouse and human embryonic stem cells (2%–4% knockin rate) through PCR-based addition of short homology arms. scCRISPR substantially lowers the bar on mouse and human transgenesis., National Institutes of Health (U.S.) (5UL1DE019581), National Institutes of Health (U.S.) (RL1DE019021), National Institutes of Health (U.S.) (1K01DK101684-01), National Institutes of Health (U.S.) (1U01HG007037), National Institutes of Health (U.S.) (5P01NS055923), Harvard Stem Cell Institute (Sternlicht Director's Fund Award)

Published
2015

3. A biomimetic extracellular matrix for cartilage tissue engineering centered on photocurable gelatin, hyaluronic acid and chondroitin sulfate

Author

Levett, P.A., Melchels, F.P.W., Schrobback, K., Hutmacher, D.W., Malda, J., Klein, T.J., Dep Gezondheidszorg Paard, CSCA TR1, and Tissue Repair

Subjects
Materials science, Light, Cell Survival, Sus scrofa, Biomedical Engineering, Cell Count, cartilage tissue engineering, Matrix (biology), Biochemistry, Biomaterials, Extracellular matrix, gelatin, chemistry.chemical_compound, Biomimetic Materials, Elastic Modulus, Hyaluronic acid, hyaluronic acid, medicine, Animals, Humans, Chondroitin sulfate, Autologous chondrocyte implantation, Cell Shape, Molecular Biology, Cells, Cultured, hydrogels, Glycosaminoglycans, Photopolymerization, Tissue Engineering, Cartilage, Chondroitin Sulfates, X-Ray Microtomography, General Medicine, Chondrogenesis, Extracellular Matrix, photopolymerisation, medicine.anatomical_structure, Gene Expression Regulation, chemistry, Self-healing hydrogels, Biophysics, 090301 Biomaterials, Biotechnology, Biomedical engineering
Abstract

The development of hydrogels tailored for cartilage tissue engineering has been a research and clinical goal for over a decade. Directing cells towards a chondrogenic phenotype and promoting new matrix formation are significant challenges that must be overcome for the successful application of hydrogels in cartilage tissue therapies. Gelatin-methacrylamide (Gel-MA) hydrogels have shown promise for the repair of some tissues, but have not been extensively investigated for cartilage tissue engineering. We encapsulated human chondrocytes in Gel-MA-based hydrogels, and show that with the incorporation of small quantities of photocrosslinkable hyaluronic acid methacrylate (HA-MA), and to a lesser extent chondroitin sulfate methacrylate (CS-MA), chondrogenesis and mechanical properties can be enhanced. The addition of HA-MA to Gel-MA constructs resulted in more rounded cell morphologies, enhanced chondrogenesis as assessed by gene expression and immunofluorescence, and increased quantity and distribution of the newly synthesized extracellular matrix (ECM) throughout the construct. Consequently, while the compressive moduli of control Gel-MA constructs increased by 26 kPa after 8 weeks culture, constructs with HA-MA and CS-MA increased by 114 kPa. The enhanced chondrogenic differentiation, distribution of ECM, and improved mechanical properties make these materials potential candidates for cartilage tissue engineering applications.

Published
2014

4. Prolonged presence of VEGF promotes vascularization in 3D bioprinted scaffolds with defined architecture

Author

Faculteit Diergeneeskunde, LS Equine Muscoskeletal Biology, Onderzoek, CSCA TR1, Tissue Repair, Poldervaart, Michelle T, Gremmels, Hendrik, van Deventer, Kelly, Fledderus, Joost O, Oner, F Cumhur, Verhaar, Marianne C, Dhert, Wouter J A, Alblas, Jacqueline, Faculteit Diergeneeskunde, LS Equine Muscoskeletal Biology, Onderzoek, CSCA TR1, Tissue Repair, Poldervaart, Michelle T, Gremmels, Hendrik, van Deventer, Kelly, Fledderus, Joost O, Oner, F Cumhur, Verhaar, Marianne C, Dhert, Wouter J A, and Alblas, Jacqueline

Published
2014

5. Prevalence and co-occurrence of hip dysplasia and elbow dysplasia in Dutch pure-bred dogs

Author

Onderzoek, LS Algemene chirurgie, Sub Neuro Chirurgie, Sub Neuro/Tandheelkunde, Tissue Repair, CSCA TR1, CSCA AVM, ES AVM, Advances in Veterinary Medicine, Lavrijsen, Ineke, Heuven, Henri, Meij, Bjorn, Theyse, Lars, Nap, R.C., Leegwater, Peter, Hazewinkel, Herman, Onderzoek, LS Algemene chirurgie, Sub Neuro Chirurgie, Sub Neuro/Tandheelkunde, Tissue Repair, CSCA TR1, CSCA AVM, ES AVM, Advances in Veterinary Medicine, Lavrijsen, Ineke, Heuven, Henri, Meij, Bjorn, Theyse, Lars, Nap, R.C., Leegwater, Peter, and Hazewinkel, Herman

Published
2014

6. In vitro induction of alkaline phosphatase levels predicts in vivo bone forming capacity of human bone marrow stromal cells.

Author

Dep Gezondheidszorg Paard, Geneeskunde van gezelschapsdieren, CSCA TR1, Tissue Repair, Prins, H.J., Braat, A.K., Gawlitta, D., Dhert, W.J.A., Egan, D.A., Tijssen-Slump, E., Yuan, H., Coffer, P.J, Rozemuller, H., Martens, A.C.M., Dep Gezondheidszorg Paard, Geneeskunde van gezelschapsdieren, CSCA TR1, Tissue Repair, Prins, H.J., Braat, A.K., Gawlitta, D., Dhert, W.J.A., Egan, D.A., Tijssen-Slump, E., Yuan, H., Coffer, P.J, Rozemuller, H., and Martens, A.C.M.

Published
2014

7. Ethical implications of regenerative medicine in orthopedics: an empirical study with surgeons and scientists in the field

Author

Faculteit Diergeneeskunde, LS Equine Muscoskeletal Biology, Onderzoek, Tissue Repair, CSCA TR1, Niemansburg, Sophie L, van Delden, Johannes J M, Oner, F Cumhur, Dhert, Wouter J A, Bredenoord, Annelien L, Faculteit Diergeneeskunde, LS Equine Muscoskeletal Biology, Onderzoek, Tissue Repair, CSCA TR1, Niemansburg, Sophie L, van Delden, Johannes J M, Oner, F Cumhur, Dhert, Wouter J A, and Bredenoord, Annelien L

Published
2014

9. Cell type and transfection reagent-dependent effects on viability, cell content, cell cycle and inflammation of RNAi in human primary mesenchymal cells

Author

Faculteit Diergeneeskunde, LS Equine Muscoskeletal Biology, Onderzoek, Tissue Repair, CSCA TR1, Yang, Hsiao-yin, Vonk, Lucienne A, Licht, Ruud, van Boxtel, Antonetta M G, Bekkers, Joris E J, Kragten, Angela H M, Hein, San, Varghese, Oommen P, Howard, Kenneth A, Öner, F Cumhur, Dhert, Wouter J A, Creemers, Laura B, Faculteit Diergeneeskunde, LS Equine Muscoskeletal Biology, Onderzoek, Tissue Repair, CSCA TR1, Yang, Hsiao-yin, Vonk, Lucienne A, Licht, Ruud, van Boxtel, Antonetta M G, Bekkers, Joris E J, Kragten, Angela H M, Hein, San, Varghese, Oommen P, Howard, Kenneth A, Öner, F Cumhur, Dhert, Wouter J A, and Creemers, Laura B

Published
2014

10. Wnt AND NOTCH REGULATE PROLIFERATION, MIGRATION AND DIFFERENTIATION OF HUMAN HEPATIC PROGENITOR CELLS AND CANINE LIVER ORGANOIDS: Journal of Hepatology

Author

Onderzoek, Sub Neuro/Tandheelkunde, CSCA TR1, LS Interne geneeskunde, Biochemisch laboratorium, Veterinair Pathologisch Diagnostisch Cnt, PB AVM, Dep Pathobiologie, LS Celbiologie-Algemeen, Sub Center for Cell Imaging, Sub Hepatologie, CSCA AVM, Schotanus, B.A., Nantasanti, S., Huch, M., Oosterhoff, L., Fieten, H., Kruitwagen, H.S., van Wolferen, M., Grinwis, G.C.M., Wubbolts, R., Favier, R.P., Penning, L.C., Vries, R.G.J., Clevers, H., Rothuizen, J., Spee, B., Onderzoek, Sub Neuro/Tandheelkunde, CSCA TR1, LS Interne geneeskunde, Biochemisch laboratorium, Veterinair Pathologisch Diagnostisch Cnt, PB AVM, Dep Pathobiologie, LS Celbiologie-Algemeen, Sub Center for Cell Imaging, Sub Hepatologie, CSCA AVM, Schotanus, B.A., Nantasanti, S., Huch, M., Oosterhoff, L., Fieten, H., Kruitwagen, H.S., van Wolferen, M., Grinwis, G.C.M., Wubbolts, R., Favier, R.P., Penning, L.C., Vries, R.G.J., Clevers, H., Rothuizen, J., and Spee, B.

Published
2014

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