Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
Curr Opin Biotechnol. 2017 Dec;48:203-209. doi: 10.1016/j.copbio.2017.06.001. Epub 2017 Jun 19.
The bacterial type II Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR Associated (Cas) systems, and in particular Streptococcus pyogenes CRISPR-Cas9, have been broadly applied to edit the genome of bacterial and eukaryotic cells. Cas9, which is an RNA-guided programmable nuclease, is a powerful tool for disrupting protein-coding genes. Cas9 cleaves target sites to generate a double-strand break (DSB) that is repaired via an error-prone repair process, leading to insertion/deletion mutations and gene knockouts. However, Cas9 can also be used to modulate genome function without gene disruption, enabling base editing, transcriptional and epigenetic reprogramming, genome imaging, cellular barcoding, genetic recording, and genetic computation.
细菌的 II 型簇状规则间隔短回文重复序列 (CRISPR)-CRISPR 相关 (Cas) 系统,特别是化脓性链球菌 CRISPR-Cas9,已被广泛应用于编辑细菌和真核细胞的基因组。Cas9 是一种 RNA 指导的可编程核酸酶,是一种强大的破坏蛋白编码基因的工具。Cas9 切割靶位点产生双链断裂 (DSB),该断裂通过易错修复过程进行修复,导致插入/缺失突变和基因敲除。然而,Cas9 也可用于在不破坏基因的情况下调节基因组功能,实现碱基编辑、转录和表观遗传重编程、基因组成像、细胞条形码、遗传记录和遗传计算。