High-efficiency multiplex genome editing of Streptomyces species using an engineered CRISPR/Cas system.
High-efficiency multiplex genome editing of Streptomyces species using an engineered CRISPR/Cas system.
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DOI：
10.1021/sb500351f
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发表时间：
2015-06-19
影响因子：
4.7
通讯作者：
Zhao, Huimin
中科院分区：
文献类型：
--
作者：
Cobb, Ryan E.;Wang, Yajie;Zhao, Huimin
放线菌，特别是链霉菌属的放线菌，仍然是天然产物及其同源生物合成途径的发现和工程化的宝贵宿主。然而，这些细菌的基因操作通常是劳动和时间密集型的。在这里，我们提出了一种用于链霉菌菌株的快速多重基因组编辑的工程CRISPR/Cas系统，证明了三种不同链霉菌物种和各种大小（范围从20 bp到30 kb）的靶向染色体缺失，效率范围从70%到100%。所设计的pCRISPomyces质粒适合分别通过Golden Gate组装和等温组装（或传统的消化/连接）组装间隔区和编辑模板，从而允许快速质粒构建以靶向任何感兴趣的基因组基因座。因此，pCRISPomyces系统代表了链霉菌基因组编辑的强大新工具。
Actinobacteria, particularly those of genus Streptomyces, remain invaluable hosts for the discovery and engineering of natural products and their cognate biosynthetic pathways. However, genetic manipulation of these bacteria is often labor and time intensive. Here, we present an engineered CRISPR/Cas system for rapid multiplex genome editing of Streptomyces strains, demonstrating targeted chromosomal deletions in three different Streptomyces species and of various sizes (ranging from 20 bp to 30 kb) with efficiency ranging from 70 to 100%. The designed pCRISPomyces plasmids are amenable to assembly of spacers and editing templates via Golden Gate assembly and isothermal assembly (or traditional digestion/ligation), respectively, allowing rapid plasmid construction to target any genomic locus of interest. As such, the pCRISPomyces system represents a powerful new tool for genome editing in Streptomyces.
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影响因子：
48
作者：
Mali, Prashant;Esvelt, Kevin M.;Church, George M.
通讯作者：
Church, George M.
影响因子：
5.7
作者：
Olano C;García I;González A;Rodriguez M;Rozas D;Rubio J;Sánchez-Hidalgo M;Braña AF;Méndez C;Salas JA
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Baltz, Richard H.
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作者：
OTTEN, SL;STUTZMANENGWALL, KJ;HUTCHINSON, CR
通讯作者：
HUTCHINSON, CR