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开发一个CRISPR/Cpf1系统用于多重基因编辑Aspergillus oryzae

Tianming Chen¹, Ziming Chen¹, Huanxin Zhang²

¹Jiangxi Key Laboratory of Bioprocess Engineering, College of Life Sciences, Jiangxi Science and Technology Normal University, Nanchang, 330013, China.

Folia microbiologica

|July 25, 2023

概括

这项研究引入了一种新的CRISPR/Cpf1系统,用于在Aspergillus oryzae中进行多重基因编辑. 弗朗西塞拉新生菌Cpf1 (FnCpf1) 系统能够同时修改多个基因,在这种真菌中推进基因组工程.

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:

Aspergillus oryzae是一种具有工业意义的细丝状真菌.
克里斯帕/卡斯技术是基因组工程的一个关键工具.
之前的工作使用了A. oryzae中的CRISPR/LbCpf1,但多重编辑尚未被探索.

研究的目的:

在A. oryzae.中开发和评估一种新的CRISPR/Cpf1多重基因编辑系统.
为了证明该系统对同时基因修饰的能力.

主要方法:

通过使用Francisella novicida Cpf1 (FnCpf1) 核酶,设计了一个CRISPR/Cpf1系统.
在A. oryzae U6促进体和终结体的控制下构建了一个CRISPR-RNA (crRNA) 表达盒,直接重复和指导序列.
设计了两个针对wa和pyrG基因的指导序列.

主要成果:

通过使用FnCpf1系统单独成功突变了wa和pyrG基因.
在A. oryzae中使用单个crRNA阵列与两个指导序列实现了wA和pyrG基因的同时编辑.
验证了开发的CRISPR/Cpf1多重系统的效率.

结论:

使用FnCpf1开发的CRISPR/Cpf1系统对A. oryzae的多重基因组编辑有效.
这个系统为A. oryzae的基因工程提供了一个强大的新工具.
同时进行基因编辑的能力提高了工业真菌复杂基因修改的潜力.

关键词:

亚斯伯吉illus oryzae 是一个植物.在CRISPR/Cpf1系统中.状真菌是一种状真菌.多重基因编辑多重基因编辑

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