Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2013-03-26

AUTHORS

Nannan Chang, Changhong Sun, Lu Gao, Dan Zhu, Xiufei Xu, Xiaojun Zhu, Jing-Wei Xiong, Jianzhong Jeff Xi

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 etsrpy11 mutant embryos or cardia bifida phenotypes in fautm236a 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. More... »

PAGES

465-472

Journal

TITLE

Cell Research

ISSUE

4

VOLUME

23

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/cr.2013.45

    DOI

    http://dx.doi.org/10.1038/cr.2013.45

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1006654084

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/23528705


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