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CRISPR/Cas9-based genome engineering in the filamentous fungus Rhizopus oryzae and its application to L-lactic acid production

文献类型：期刊论文

作者Zhu, Haodong1,2; Wang, Han2; Wang, Li2; Zheng, Zhiming1,2
刊名BIOTECHNOLOGY JOURNAL
出版日期2024-09-01
卷号19
关键词CRISPR/Cas9 system genome editing Rhizopus oryzae U6 promoter
ISSN号1860-6768
DOI10.1002/biot.202400309
通讯作者Wang, Han(wanghan@ipp.ac.cn) ; Zheng, Zhiming(zhengzhiming2014@163.com)
英文摘要The filamentous fungus Rhizopus oryzae is one of the main industrial strains for the production of a series of important chemicals such as ethanol, lactic acid, and fumaric acid. However, the lack of efficient gene editing tools suitable for R. oryzae makes it difficult to apply technical methods such as metabolic engineering regulation and synthetic biology modification. A CRISPR-Cas9 system suitable for efficient genome editing in R. oryzae was developed. Firstly, four endogenous U6 promoters of R. oryzae were identified and screened with the highest transcriptional activity for application to sgRNA transcription. It was then determined that the U6 promoter mediated CRISPR/Cas9 system has the ability to efficiently edit the genome of R. oryzae through NHEJ and HDR-mediated events. Furthermore, the newly constructed CRISPR-Cas9 dual sgRNAs system can simultaneously disrupt or insert different fragments of the R. oryzae genome. Finally, this CRISPR-Cas9 system was applied to the genome editing of R. oryzae by knocking out pyruvate carboxylase gene (PYC) and pyruvate decarboxylase gene (pdcA) and knocking in phosphofructokinase (pfkB) from Escherichia coli and L-lactate dehydrogenase (L-LDH) from Heyndrickxia coagulans, which resulted in a substantial increase in L-LA production. In summary, this study showed that the CRISPR/Cas9-based genome editing tool is efficient for manipulating genes in R. oryzae.
WOS关键词SGRNA DESIGN ; FERMENTATION ; GENE
资助项目Anhui Province Key Research and Development Plan ; National Key Research and Development Program of China[2019YFA0904304] ; Hefei Institutes of Physical Science, Chinese Academy of Sciences Director's Fund[YZJJ2024QN37] ; [S2023n06020216]
WOS研究方向Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种英语
WOS记录号WOS:001315671000001
出版者WILEY-V C H VERLAG GMBH
资助机构Anhui Province Key Research and Development Plan ; National Key Research and Development Program of China ; Hefei Institutes of Physical Science, Chinese Academy of Sciences Director's Fund
源URL[http://ir.hfcas.ac.cn:8080/handle/334002/135380]  
专题中国科学院合肥物质科学研究院
通讯作者Wang, Han; Zheng, Zhiming
作者单位1.Univ Sci & Technol China, Sci Isl Branch Grad, Hefei, Anhui, Peoples R China
2.Chinese Acad Sci, Hefei Inst Phys Sci, POB 1138,Comprehens Lab Bldg,350 Shushanhu Rd, Hefei 230031, Anhui, Peoples R China
推荐引用方式
GB/T 7714		 Zhu, Haodong,Wang, Han,Wang, Li,et al. CRISPR/Cas9-based genome engineering in the filamentous fungus Rhizopus oryzae and its application to L-lactic acid production[J]. BIOTECHNOLOGY JOURNAL,2024,19.
APA Zhu, Haodong,Wang, Han,Wang, Li,&Zheng, Zhiming.(2024).CRISPR/Cas9-based genome engineering in the filamentous fungus Rhizopus oryzae and its application to L-lactic acid production.BIOTECHNOLOGY JOURNAL,19.
MLA Zhu, Haodong,et al."CRISPR/Cas9-based genome engineering in the filamentous fungus Rhizopus oryzae and its application to L-lactic acid production".BIOTECHNOLOGY JOURNAL 19(2024).

入库方式： OAI收割

来源：合肥物质科学研究院

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