同源重组
大肠杆菌
素数(序理论)
DNA
计算生物学
记录
转座因子
合成生物学
核酸外切酶
核酸外切酶 III
基因组编辑
生物
突变体
遗传学
基因组
DNA修复
基因
DNA聚合酶
组合数学
数学
作者
Hongyuan Zhang,Jiacheng Ma,Zhaowei Wu,Xiaoyang Chen,Qian Yin,Weizhong Chen,Z Wang,Ya Ping Zhang,Huanhu Zhu,Xingxu Huang,Quanjiang Ji
标识
DOI:10.1038/s41467-024-45114-4
摘要
Prime editing allows precise installation of any single base substitution and small insertions and deletions without requiring homologous recombination or double-strand DNA breaks in eukaryotic cells. However, the applications in bacteria are hindered and the underlying mechanisms that impede efficient prime editing remain enigmatic. Here, we report the determination of vital cellular factors that affect prime editing in bacteria. Genetic screening of 129 Escherichia coli transposon mutants identified sbcB, a 3'→5' DNA exonuclease, as a key genetic determinant in impeding prime editing in E. coli, combinational deletions of which with two additional 3'→5' DNA exonucleases, xseA and exoX, drastically enhanced the prime editing efficiency by up to 100-fold. Efficient prime editing in wild-type E. coli can be achieved by simultaneously inhibiting the DNA exonucleases via CRISPRi. Our results pave the way for versatile applications of prime editing for bacterial genome engineering.
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