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工程 Pichia pastoris 菌株使用CRISPR/Cas9技术:基本协议

Hana Raschmanová¹, Astrid Weninger², Karin Kovar³

¹University of Chemistry and Technology Prague, Department of Biotechnology, Prague, Czech Republic.

Methods in molecular biology (Clifton, N.J.)

|October 1, 2025

概括

这项研究详细介绍了Pichia pastoris的强大的CRISPR/Cas9基因组编辑协议,在基因失活方面实现了近100%的效率. 这种方法促进了酵母的基因修改和代谢工程.

科学领域:

分子生物学分子生物学
酵母遗传学酵母遗传学

背景情况:

克里斯普尔/卡斯9系统是基因组编辑和代谢工程的强大工具.
在2016年建立的,CRISPR/Cas9协议用于Komagataella phaffii (Pichia pastoris) 允许基因修饰,如无标记基因破坏.
以前的应用包括提高同源重组效率.

研究的目的:

描述一个强大的,基于CRISPR的基因组编辑在Pichia pastoris的基本协议.
通过使用该协议,证明基因失活的高准效率.

主要方法:

利用CRISPR/Cas9系统在Pichia pastoris中进行基因组编辑.
为了基因失活,采用了一个框架转移突变策略.
建立了P. pastoris基因组编辑的基本,强大的协议.

主要成果:

在基因失活方面实现了接近100%的准效率.
证明了基于CRISPR的基因改造协议的有效性.
该协议适合创建用于基因淘汰的框架转移突变.

结论:

描述的CRISPR/Cas9协议为Pichia pastoris基因组编辑提供了一种可靠的方法.
该协议有助于有效的基因失活和其他遗传修饰.
对P. pastoris的CRISPR/Cas9技术的进一步优化可用于特定的应用.

关键词:

这就是CRISPR/Cas9的作用.基因淘汰赛基因淘汰赛基因组编辑基因组编辑基因组工程是基因组工程.指南RNA指南RNA是指导RNA的指南RNA.同类的重组组合.科玛格塔贝拉 (Komagataella phaffii) 是一个有趣的植物.这就是Pichia pastoris.合成生物学合成生物学

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