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在Aspergillus niger中使用多重化CRISPRoff系统对FlbA调节的转录因子基因的功能分析

Xiaoyi Chen¹, Juan P Moran Torres¹, S Vyanjan K Tedjai¹

¹Microbiology, Department of Biology, Utrecht University, Padualaan 8, 3584, CH, Utrecht, the Netherlands.

International journal of biological macromolecules

|August 1, 2024

概括

研究人员使用CRISPRoff研究阿斯伯吉勒斯尼日尔的转录因子,识别了像dofA和soca这样的基因,这些基因参与了胞,生长和酶分泌. 这突出了CRISPR的功能.

科学领域:

菌遗传学和分子生物学
基因调节和表观遗传学
亚斯伯菌尼格尔研究研究

背景情况:

尼日尔菌中的FlbA调节了36个转录因子 (TF) 基因,影响分泌,生长和分泌.
了解这些TF基因的特定作用对于真菌发育至关重要.

研究的目的:

为了功能性地分析Aspergillus niger中FlbA调节的TF基因的一个子集.
评估CRISPRoff系统在真菌基因功能研究中的有效性.

主要方法:

采用了优化的CRISPRoff系统和一套sgRNA图书馆,针对36个TF基因的促销者.
引入了CRISPRoff等离子体进入黑色阿斯伯吉路斯.
选的转化剂用于分泌,生物质和分泌表型.

主要成果:

确定了五个关键的TF基因 (dofA,dofB,dofC,dofD,soca) 影响胞和细胞外酶活性.
还发现dofA和soca也参与生物质形成.
在真菌基因功能分析中证明了多重化CRISPRoff的成功应用.

结论:

CRISPRoff系统是Aspergillus niger多重功能基因组学的一个有效工具.
确定了在真菌发育和生理学中发挥重要作用的特定转录因子 (dofA,dofB,dofC,dofD,soca).

关键词:

亚斯珀吉洛斯 (Aspergillus) 是一种植物.表观遗传修饰是一种表观遗传修饰.这种真菌是真菌.

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相关概念视频

CRISPR and crRNAs

CRISPR and crRNAs

Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...