清脆的
计算生物学
Cas9
质粒
基因组
DNA
多路复用
生物
基因
遗传学
突变
作者
Hee‐Jung Lim,Soyeong Jun,Minjeong Park,Junghak Lim,Jaehwan Jeong,Ji Hyun Lee,Duhee Bang
标识
DOI:10.1021/acssynbio.0c00002
摘要
We developed a clustered regularly interspaced short palindromic repeats (CRISPR)/retron system for multiplexed generation of substitution mutations by coutilization of a retron system that continuously expresses donor DNA and a CRISPR/Cas9 cassette that induces cleavage at target genomic loci. Our system efficiently introduces substitution mutation in the Escherichia coli genome in a high-throughput manner. These substitution mutations can be tracked by analysis of retron plasmid sequences without laborious amplification of individual edited loci. We demonstrated that our CRISPR/retron system can introduce thousands of mutations in a single experiment and be used for screening phenotypes related to chemical responses or fitness changes. We expect that our system could facilitate genome-scale substitution screenings.
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