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利用源自Streptomyces的I-E型CRISPR/Cas系统在Streptomyces中进行高效的基因组编辑

Yuhui Xie¹, Xiaoyan Liu¹, Tingting Wu²

¹Frontiers 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.

Science China. Life sciences

|January 17, 2025

概括

科学家们设计了一个新的CRISPR/Cas基因编辑工具,用于Streptomyces细菌. 这个系统有效地修改DNA,帮助在这些重要的微生物中发现天然产品.

科学领域:

微生物学微生物学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

克里斯普尔/卡斯系统是微生物研究和自然产品开发的强大遗传工具.
2类CRISPR/Cas系统被广泛使用,但在Streptomyces中通常是无效的.
在生产有价值的自然产品方面,Streptomyces的作用至关重要.

研究的目的:

开发一种高效的基因编辑系统,用于各种Streptomyces菌株.
克服现有的CRISPR/Cas工具在Streptomyces中的局限性.
为了提高从Streptomyces天然产品的发现.

主要方法:

设计了一种来自Streptomyces avermitilis的第1类I型CRISPR/Cas系统.
使用等离子体干扰试验确定了原空间体相邻的动图 (5'-AAN-3').
应用了针对性染色体删除和DNA插入的系统.

主要成果:

在基因学上遥远的Streptomyces菌株中实现了高效的基因编辑.
成功执行了从8bp到100kb的染色体删除,效率>92%.
促进异质基因表达和自然产物基因集群的激活.

结论:

建立了一种基于CRISPR/Cas的新型I型Streptomyces基因编辑方法.
这是第一个基因编辑工具,基于Streptomyces内源性1类I型CRISPR/Cas系统.
开发的系统扩大了Streptomyces的基因操纵工具箱,推动了天然产品的发现.

关键词:

这种病名为Streptomyces.基因组编辑基因组编辑标识标识标识标识标识自然产品是一种自然产品.原太空体相邻动图 (PAM)类型 I-E CRISPR/Cas 系统

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