Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1, Yayoi, Tokyo 113-8657, Japan.
Int J Mol Sci. 2017 Oct 31;18(11):2286. doi: 10.3390/ijms18112286.
Mammalian zygote-mediated genome-engineering by Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas is currently used for the generation of genome-modified animals. Here, we report that a -derived orthologous CRISPR/Cas system recognizes a 5'-NNNVRYAC sequence as a protospacer-adjacent motif in mouse zygotes, and is applicable for efficient generation of knockout mice. Moreover, this novel CRISPR/Cas can be used for zygote-mediated knock-in at a unique locus, suggesting that this system could help to expand the feasibility of the zygote-mediated generation of genome-modified animals.
哺乳动物受精卵介导的基因组工程通过成簇规律间隔短回文重复序列 (CRISPR)/Cas 目前用于产生基因组修饰动物。在这里,我们报告说,一种 - 衍生的同源 CRISPR/Cas 系统在小鼠受精卵中识别 5'-NNNVRYAC 序列作为前导间隔基序,并且可用于有效产生基因敲除小鼠。此外,这种新型 CRISPR/Cas 可用于在独特基因座进行受精卵介导的基因敲入,表明该系统有助于扩大受精卵介导的基因组修饰动物产生的可行性。