利用嗜热链球菌1来源的CRISPR/Cas系统通过合子介导产生基因组修饰小鼠。
Zygote-mediated generation of genome-modified mice using Streptococcus thermophilus 1-derived CRISPR/Cas system.
作者信息
Fujii Wataru, Kakuta Shigeru, Yoshioka Shin, Kyuwa Shigeru, Sugiura Koji, Naito Kunihiko
机构信息
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan.
Department of Biomedical Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan.
出版信息
Biochem Biophys Res Commun. 2016 Aug 26;477(3):473-6. doi: 10.1016/j.bbrc.2016.06.070. Epub 2016 Jun 16.
Mammalian zygote-mediated genome-engineering by CRISPR/Cas is currently used for the generation of genome-modified animals. Here we report that a Streptococcus thermophilus-1 derived orthologous CRISPR/Cas system, which recognizes the 5'-NNAGAA sequence as a protospacer adjacent motif (PAM), is useful in mouse zygotes and is applicable for generating knockout mice (87.5%) and targeted knock-in mice (45.5%). The induced mutation could be inherited in the next generation. This novel CRISPR/Cas can expand the feasibility of the zygote-mediated generation of genome-modified animals that require an exact mutation design.
目前，通过CRISPR/Cas进行的哺乳动物合子介导的基因组工程被用于生成基因组修饰动物。在此，我们报告一种源自嗜热链球菌-1的直系同源CRISPR/Cas系统，该系统将5'-NNAGAA序列识别为原间隔相邻基序（PAM），在小鼠合子中有效，可用于生成基因敲除小鼠（87.5%）和靶向敲入小鼠（45.5%）。诱导的突变能够在下一代中遗传。这种新型的CRISPR/Cas可以扩展合子介导的基因组修饰动物生成的可行性，而这需要精确的突变设计。
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