Suppr超能文献

一种用于最小化 DNA 碱基编辑器脱靶效应的 Cas 嵌入策略。

A Cas-embedding strategy for minimizing off-target effects of DNA base editors.

机构信息

School of Life Science and Technology, ShanghaiTech University, Shanghai, China.

Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.

出版信息

Nat Commun. 2020 Nov 27;11(1):6073. doi: 10.1038/s41467-020-19690-0.

Abstract

DNA base editors, typically comprising editing enzymes fused to the N-terminus of nCas9, display off-target effects on DNA and/or RNA, which have remained an obstacle to their clinical applications. Off-target edits are typically countered via rationally designed point mutations, but the approach is tedious and not always effective. Here, we report that the off-target effects of both A > G and C > T editors can be dramatically reduced without compromising the on-target editing simply by inserting the editing enzymes into the middle of nCas9 at tolerant sites identified using a transposon-based genetic screen. Furthermore, employing this Cas-embedding strategy, we have created a highly specific editor capable of efficient C > T editing at methylated and GC-rich sequences.

摘要

DNA 碱基编辑器通常由与 nCas9 N 端融合的编辑酶组成,在 DNA 和/或 RNA 上显示出脱靶效应,这一直是其临床应用的障碍。通常通过合理设计的点突变来对抗脱靶编辑,但这种方法繁琐且并非总是有效。在这里,我们报告说,通过在基于转座子的遗传筛选中鉴定的耐受位点将编辑酶插入 nCas9 的中间,可以在不影响靶编辑的情况下,显著降低 A>G 和 C>T 编辑器的脱靶效应。此外,采用这种 Cas 嵌入策略,我们创建了一种高度特异性的编辑器,能够在甲基化和富含 GC 的序列中有效进行 C>T 编辑。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9a69/7695861/cb789f815d24/41467_2020_19690_Fig1_HTML.jpg

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验