Harnessing the Streptomyces-originating typeⅠ-E CRISPR/Cas system for efficient genome editing in Streptomyces  

作　　者：Yuhui Xie Xiaoyan Liu Tingting Wu Yunzi Luo 

机构地区：[1]Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering(Ministry of Education),School of Chemical Engineering and Technology,Tianjin University,Tianjin,300072,China [2]Georgia Tech Shenzhen Institute,Tianjin University,Shenzhen,518071,China [3]Haihe Laboratory of Sustainable Chemical Transformations,Tianjin,300192,China [4]Department of Gastroenterology,State Key Laboratory of Biotherapy,West China Hospital,Sichuan University,Chengdu,610041,China

出　　处：《Science China(Life Sciences)》2025年第4期1174-1182,共9页中国科学(生命科学英文版)

基　　金：supported by the National Key Research and Development Program of China(2018YFA0903300);the National Natural Science Foundation of China(32071426);the KeyArea Research and Development Program of Guangdong Province(2020B0303070002);the Haihe Laboratory of Sustainable Chemical Transformations(24HHWCSS00006)。

摘　　要：Since their discovery,CRISPR/Cas systems have significantly expanded the genetic toolbox,aiding in the exploration and enhanced production of natural products across various microbes.Among these,class 2 CRISPR/Cas systems are simpler and more broadly used,but they frequently fail to function effectively in many Streptomyces strains.In this study,we present an engineered class 1 typeⅠCRISPR/Cas system derived from Streptomyces avermitilis,which enables efficient gene editing in phylogenetically distant Streptomyces strains.Through a plasmid interference assay,we identified the effective protospacer adjacent motif as 5′-AAN-3′.Utilizing this system,we achieved targeted chromosomal deletions ranging from 8 bp to 100 kb,with efficiencies exceeding 92%.We further utilized this system to insert DNA fragments into different Streptomyces genomes,facilitating the heterologous expression of exogenous genes and the activation of endogenous natural product biosynthetic gene clusters.Overall,we established a typeⅠCRISPR/Cas-based gene-editing methodology that significantly advances the exploration of Streptomyces,known for their rich natural product resources.This is the first report of a gene editing tool developed based on the endogenous class 1 typeⅠCRISPR/Cas system in Streptomyces spp.Our work enriches the Streptomyces gene manipulation toolbox and advances the discovery of valuable natural products within these organisms.

关 键 词：genome editing type I-E CRISPR/Cas system STREPTOMYCES protospacer adjacent motif(PAM) identification natural product 

分 类 号：Q78[生物学—分子生物学]