Efficient generation of selection‐gene‐free rat knockout models by homologous recombination in ES cells

Embryonic stem cell (ES cell)‐based rat knockout technology, although successfully developed in 2010, has seen very limited usage to date due to low targeting efficiency and a lack of optimized procedures. In this study, we performed gene targeting in ES cells from the Sprague–Dawley (SD) and the Fi...

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Autores principales: Lan, He, Li, Shuping, Guo, Zihang, Men, Hongsheng, Wu, Yuanyuan, Li, Ning, Bryda, Elizabeth C., Capecchi, Mario R., Wu, Sen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2016
Materias:
Research Letters
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5129459/
https://www.ncbi.nlm.nih.gov/pubmed/27597178
http://dx.doi.org/10.1002/1873-3468.12388
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author Lan, He
Li, Shuping
Guo, Zihang
Men, Hongsheng
Wu, Yuanyuan
Li, Ning
Bryda, Elizabeth C.
Capecchi, Mario R.
Wu, Sen
author_facet Lan, He
Li, Shuping
Guo, Zihang
Men, Hongsheng
Wu, Yuanyuan
Li, Ning
Bryda, Elizabeth C.
Capecchi, Mario R.
Wu, Sen
author_sort Lan, He
collection PubMed
description Embryonic stem cell (ES cell)‐based rat knockout technology, although successfully developed in 2010, has seen very limited usage to date due to low targeting efficiency and a lack of optimized procedures. In this study, we performed gene targeting in ES cells from the Sprague–Dawley (SD) and the Fischer 344 (F344) rat strains using an optimized procedure and the self‐excising neomycin (neo)‐positive selection cassette ACN to successfully generate Leptin and Trp53 knockout rats that did not carry the selection gene. These results demonstrate that our simplified targeting strategy using ACN provides an efficient approach to knock out many other rat genes.
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spelling pubmed-51294592016-11-30 Efficient generation of selection‐gene‐free rat knockout models by homologous recombination in ES cells Lan, He Li, Shuping Guo, Zihang Men, Hongsheng Wu, Yuanyuan Li, Ning Bryda, Elizabeth C. Capecchi, Mario R. Wu, Sen FEBS Lett Research Letters Embryonic stem cell (ES cell)‐based rat knockout technology, although successfully developed in 2010, has seen very limited usage to date due to low targeting efficiency and a lack of optimized procedures. In this study, we performed gene targeting in ES cells from the Sprague–Dawley (SD) and the Fischer 344 (F344) rat strains using an optimized procedure and the self‐excising neomycin (neo)‐positive selection cassette ACN to successfully generate Leptin and Trp53 knockout rats that did not carry the selection gene. These results demonstrate that our simplified targeting strategy using ACN provides an efficient approach to knock out many other rat genes. John Wiley and Sons Inc. 2016-09-18 2016-10 /pmc/articles/PMC5129459/ /pubmed/27597178 http://dx.doi.org/10.1002/1873-3468.12388 Text en © 2016 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial (http://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Research Letters
Lan, He
Li, Shuping
Guo, Zihang
Men, Hongsheng
Wu, Yuanyuan
Li, Ning
Bryda, Elizabeth C.
Capecchi, Mario R.
Wu, Sen
Efficient generation of selection‐gene‐free rat knockout models by homologous recombination in ES cells
title Efficient generation of selection‐gene‐free rat knockout models by homologous recombination in ES cells
title_full Efficient generation of selection‐gene‐free rat knockout models by homologous recombination in ES cells
title_fullStr Efficient generation of selection‐gene‐free rat knockout models by homologous recombination in ES cells
title_full_unstemmed Efficient generation of selection‐gene‐free rat knockout models by homologous recombination in ES cells
title_short Efficient generation of selection‐gene‐free rat knockout models by homologous recombination in ES cells
title_sort efficient generation of selection‐gene‐free rat knockout models by homologous recombination in es cells
topic Research Letters
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5129459/
https://www.ncbi.nlm.nih.gov/pubmed/27597178
http://dx.doi.org/10.1002/1873-3468.12388
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