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Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos
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Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos
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Journal Article
Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos
2013
Overview
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 etsrpv11 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, envi- ronmental studies and medicine.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ Animals
/ Biomedical and Life Sciences
/ Embryos
/ GATA Transcription Factors - genetics
/ GATA Transcription Factors - metabolism
/ GATA5 Transcription Factor - genetics
/ GATA5 Transcription Factor - metabolism
/ Genome
/ Mammals
/ Mutants
/ Mutation
/ Original
/ RNA
/ Zebrafish Proteins - genetics
/ Zebrafish Proteins - metabolism
/ 体细胞突变
/ 制导
/ 基因组
/ 斑马鱼
/ 核酸
/ 编辑
/ 胚胎
