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Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos

Open access
Chang, Nannan, Sun, Changhong, Gao, Lu, Zhu, Dan, Xu, Xiufei, Zhu, Xiaojun, Xiong, Jing-Wei, Xi, Jianzhong Jeff·2013·Cell Research

Abstract

Recent advances with the type II clustered regularly interspaced short palindromic repeats (CRISPR) system promise an improved approach to genome editing. However, the applicability and efficiency of this system in model organisms, such as zebrafish, are little studied. Here, we report that RNA-guided Cas9 nuclease efficiently facilitates genome editing in both mammalian cells and zebrafish embryos in a simple and robust manner. Over 35% of site-specific somatic mutations were found when specific Cas/gRNA was used to target either etsrp, gata4 or gata5 in zebrafish embryos in vivo. The Cas9/gRNA efficiently induced biallelic conversion of etsrp or gata5 in the resulting somatic cells, recapitulating their respective vessel phenotypes in etsrp(y11) mutant embryos or cardia bifida phenotypes in fau(tm236a) mutant embryos. Finally, we successfully achieved site-specific insertion of mloxP sequence induced by Cas9/gRNA system in zebrafish embryos. These results demonstrate that the Cas9/gRNA system has the potential of becoming a simple, robust and efficient reverse genetic tool for zebrafish and other model organisms. Together with other genome-engineering technologies, the Cas9 system is promising for applications in biology, agriculture, environmental studies and medicine.

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Metadata

Authors
Chang, N., Sun, C., Gao, L., Zhu, D., Xu, X., Zhu, X., Xiong, J.-W., & Xi, J. J.
Venue
Cell Research
Published
2013-03-26
Volume
23
Issue
4
Pages
465-472
ISSN
  • 1001-0602 (ISSN-L)
  • 1748-7838
OpenAlex
W2001497035
Cited by
820
References
6
FWCI
41.96
Type
journal article
Language
en
Open access
hybrid

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CoChat. (2026). Citation check: Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos. https://app.cochat.ai/papers/10.1038/cr.2013.45
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Chang, N., Sun, C., Gao, L., Zhu, D., Xu, X., Zhu, X., Xiong, J.-W., & Xi, J. J. (2013). Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos. Cell Research. https://doi.org/10.1038/cr.2013.45

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