清脆的
转座因子
流动遗传元素
生物
转座酶
计算生物学
基因组编辑
整合酶
基因组
DNA
遗传学
基因
质粒
作者
Sanne E. Klompe,Phuc Leo H. Vo,Tyler S. Halpin-Healy,Samuel H. Sternberg
出处
期刊:Nature
[Springer Nature]
日期:2019-06-12
卷期号:571 (7764): 219-225
被引量:477
标识
DOI:10.1038/s41586-019-1323-z
摘要
Conventional CRISPR-Cas systems maintain genomic integrity by leveraging guide RNAs for the nuclease-dependent degradation of mobile genetic elements, including plasmids and viruses. Here we describe a notable inversion of this paradigm, in which bacterial Tn7-like transposons have co-opted nuclease-deficient CRISPR-Cas systems to catalyse RNA-guided integration of mobile genetic elements into the genome. Programmable transposition of Vibrio cholerae Tn6677 in Escherichia coli requires CRISPR- and transposon-associated molecular machineries, including a co-complex between the DNA-targeting complex Cascade and the transposition protein TniQ. Integration of donor DNA occurs in one of two possible orientations at a fixed distance downstream of target DNA sequences, and can accommodate variable length genetic payloads. Deep-sequencing experiments reveal highly specific, genome-wide DNA insertion across dozens of unique target sites. This discovery of a fully programmable, RNA-guided integrase lays the foundation for genomic manipulations that obviate the requirements for double-strand breaks and homology-directed repair.
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