文献类型： 外文期刊
作者： Chen, Tianming 1 ; Chen, Ziming 1 ; Zhang, Huanxin 2 ; Li, Yuzhen 1 ; Yao, Lihua 1 ; Zeng, Bin 1 ; Zhang, Zhe 1 ;
作者机构： 1.Jiangxi Sci & Technol Normal Univ, Coll Life Sci, Jiangxi Key Lab Bioproc Engn, Nanchang 330013, Peoples R China
2.Jiangxi Acad Agr Sci, Inst Hort, Nanchang 330200, Peoples R China
3.Shenzhen Technol Univ, Coll Pharm, Shenzhen 518118, Peoples R China
关键词： CRISPR; Cpf1 system; Multiplex gene editing; Filamentous fungi; Aspergillus oryzae
期刊名称：FOLIA MICROBIOLOGICA （ 影响因子：2.6； 五年影响因子：2.6 ）
ISSN： 0015-5632
年卷期： 2023 年
页码：
收录情况： SCI
摘要： CRISPR/Cas technology is a powerful tool for genome engineering in Aspergillus oryzae as an industrially important filamentous fungus. Previous study has reported the application of the CRISPR/Cpf1 system based on the Cpf1 (LbCpf1) from Lachnospiraceae bacterium in A. oryzae. However, multiplex gene editing have not been investigated using this system. Here, we presented a new CRISPR/Cpf1 multiplex gene editing system in A. oryzae, which contains the Cpf1 nuclease (FnCpf1) from Francisella tularensis subsp. novicida U112 and CRISPR-RNA expression cassette. The crRNA cassette consisted of direct repeats and guide sequences driven by the A. oryzae U6 promoter and U6 terminator. Using the constructed FnCpf1 gene editing system, the wA and pyrG genes were mutated successfully. Furthermore, simultaneous editing of wA and pyrG genes in A. oryzae was performed using two guide sequences targeting these gene loci in a single crRNA array. This promising CRISPR/Cpf1 genome-editing system provides a powerful tool for genetically engineering A. oryzae.
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