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一个具有重组能力的同类Yarrowia lipolytica底盘用于多重基因组操纵

Lu Shan¹, Kevin J Verstrepen², Qinhong Wang³

¹State Key Laboratory of Engineering Biology for Low-carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China; University of Chinese Academy of Sciences, Beijing 100049, China; National Center of Technology Innovation for Synthetic Biology, Tianjin 300308, China; TIB - VIB Joint Center of Synthetic Biology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.

Trends in biotechnology

|July 9, 2025

概括

研究人员对Yarrowia lipolytica进行了工程设计,以支持同源重组 (HR) 与非同源末端连接 (NHEJ) 进行精确的基因组编辑. 这提高了HR的效率,使这一重要的微生物能够进行复杂的遗传修饰.

科学领域:

微生物学微生物学
分子生物学分子生物学
合成生物学合成生物学

背景情况:

同类重组 (HR) 对于精确的基因组编辑至关重要,但在许多生物体中,容易出现错误的非同类末端连接 (NHEJ) 往往会超越它.
雅罗维亚脂质菌是一种重要的工业微生物,但其基因组编辑能力受到偏好NHEJ的限制.

研究的目的:

为精确和高效的基因组编辑设计一种在同源重组 (HR) 中熟练的Yarrowia lipolytica菌株.
提高复杂的基因改造的效率,包括多片段和多站点的整合.

主要方法:

在Yarrowia lipolytica的重组机器的系统工程.
调节多次入侵诱导的重新排列 (MIR) 过程.
亲属单链DNA化蛋白 (SSAP) -单链DNA结合蛋白 (SSB) 对的表达.

主要成果:

与NHEJ缺陷菌株相比,多片段多站点集成的HR效率提高了38.9倍,其他集成类型的HR效率提高了1.6-1.2倍.
通过短50个基数对的同质手臂实现了58%的HR效率.
在两个基因组部位同时成功集成了两个大型DNA片段 (18.0kb和13.5kb).
证明了同时编辑,抑制和激活多个基因,从而增加了细胞的强度.

结论:

开发了一种新的HR熟练的Yarrowia lipolytica菌株.
这种工程菌株促进了高效和精确的基因组编辑,包括复杂的多基因和大片段集成.
为在Yarrowia lipolytica中推进基因工程应用提供了有价值的底盘.

关键词:

DSB维修服务提供者雅罗维亚 (Yarrowia lipolytica) 是一个高脂的植物.底盘压力是底盘的压力.基因工程是基因工程,是基因工程.同类的重组组合.多个基因组操纵多个基因组操纵.不同类型的末端连接.

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