Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects
Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects
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DOI：
10.1038/nmeth.2857
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发表时间：
2014-04-01
期刊：
影响因子：
48
通讯作者：
Skarnes, William C.
中科院分区：
文献类型：
--
作者：
Shen, Bin;Zhang, Wensheng;Skarnes, William C.
细菌RNA指导的Cas9内切核酸酶是用于真核生物中位点特异性基因组修饰的通用工具。用Cas9 mRNA和单向导RNA共显微注射小鼠胚胎诱导可传递给后代的中靶和脱靶突变。然而，Cas9切口酶可以用于有效地突变基因，而在已知的脱靶位点没有可检测的损伤。该方法适用于任何模式生物的基因组编辑，并最大限度地减少脱靶突变的混淆问题。
Bacterial RNA-directed Cas9 endonuclease is a versatile tool for site-specific genome modification in eukaryotes. Co-microinjection of mouse embryos with Cas9 mRNA and single guide RNAs induces on-target and off-target mutations that are transmissible to offspring. However, Cas9 nickase can be used to efficiently mutate genes without detectable damage at known off-target sites. This method is applicable for genome editing of any model organism and minimizes confounding problems of off-target mutations.