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通过使用限制修改系统静音CRISPR-Cas9工具包对大多数黄金葡萄球菌进行高效的基因组编辑

Ping Yang^1,2, Junjie Yang³, Ting Lin¹

¹School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.

ACS synthetic biology

|October 13, 2023

概括

一个新的CRISPR-Cas9工具包克服了Staphylococcus aureus中的限制障碍,使得有效的基因操纵成为可能. 这种工具加速了突变体的构建,用于研究这种重要病原体的耐药性和病原性.

科学领域:

微生物学微生物学
分子生物学分子生物学
基因工程是一种基因工程.

背景情况:

黄金葡萄球菌 (Staphylococcus aureus) 是一个重要的病原体,会引起严重的感染.
基因操纵对于了解黄金菌的病原性和耐药性至关重要.
限制障碍阻碍了在临床S. aureus分离物中使用遗传工具.

研究的目的:

开发一种新的遗传工具包,以克服S. aureus.中的限制障碍.
提高基因操纵在包括临床分离物在内的多种S. aureus菌株的效率.

主要方法:

构建一个限制修改 (RM) 系统静音的CRISPR-Cas9工具包.
从等离子体中消除I型RM目标,并使用小圆技术进行等离子体缩小.
使用等离子体人工修饰 (PAM) 方法绕过II型RM系统.

主要成果:

显著提高了转换效率 (10^5-10^6变压剂/μg等离子体).
基因删除的高成功效率,在一轮电穿孔中产生淘汰菌株.
在包括临床分离物种在内的广泛的金黄色细菌物种中证明了有效性.

结论:

无声RM的CRISPR-Cas9工具包显著提高了S. aureus的遗传操纵.
这种方法加快了突变构造,帮助研究药物耐药性和病原性.
该工具包的设计原则可用于开发更广泛的细菌应用的遗传工具.

关键词:

这就是CRISPR-Cas9的特征.黄金葡萄球菌黄金葡萄球菌基因组编辑基因组编辑限制-修改-限制的修改合成DNA是一种合成DNA.转化效率的转化效率是什么

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