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Developing a CRISPR/Cas9 System for Genome Editing in the Basidiomycetous Yeast Rhodosporidium toruloides

文献类型：期刊论文

作者Jiao, Xiang1,2; Zhang, Yue1,2; Liu, Xiangjian2; Zhang, Qi1,2; Zhang, Sufang2; Zhao, Zongbao K.2
刊名BIOTECHNOLOGY JOURNAL
出版日期2019-07-01
卷号14期号:7页码:7
ISSN号1860-6768
关键词basidiomycetous yeasts CRISPR Cas9 genome editing Rhodosporidium toruloides transformation
DOI10.1002/biot.201900036
通讯作者Zhang, Sufang(zsfjxy@dicp.ac.cn) ; Zhao, Zongbao K.(zhaozb@dicp.ac.cn)
英文摘要The basidiomycetous yeast Rhodosporidium toruloides (R. toruloides) has been explored as a promising host for the production of lipids and carotenoids. However, the rational manipulation of this yeast remains difficult due to lack of efficient genetic tools. Here, the development of a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease (Cas9) system for genome editing in R. toruloides is described. First, R. toruloides strains are generated with sufficient production of the Cas9 protein of Staphylococcus aureus origin by integrating a cassette containing a codon-optimized Cas9 gene into the genome. In parallel, two U6 genes are identified, predicting two U6 promoters and confirming better transcription of single-guide RNA (sgRNA) with the U6b promoter. Next, sgRNA cassettes are designed targeting CRTI, CAR2, and CLYBL gene, respectively, transforming into those Cas9-expressed strains, and finding over 60% transformants with successful insertion and deletion (indel) mutations. Furthermore, when the sgRNA cassette includes donor DNA flanked by two homologous arms of the gene CRTI, gene knockout occurs via homologous recombination. Thus, the CRISPR/Cas9 system is now established as a powerful genome-editing tool in R. toruloides, which should facilitate functional genomic study and advanced cell factory development.
WOS关键词OLEAGINOUS YEAST ; IN-VIVO ; GENES ; CAS9 ; TRANSFORMATION ; PROMOTERS
资助项目National Natural Science Foundation of China[31870042] ; National Natural Science Foundation of China[21721004] ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences[DMTO201701]
WOS研究方向Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种英语
出版者WILEY-V C H VERLAG GMBH
WOS记录号WOS:000476825900015
资助机构National Natural Science Foundation of China ; National Natural Science Foundation of China ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences ; Dalian Institute of Chemical Physics, Chinese Academy of Sciences
源URL[http://cas-ir.dicp.ac.cn/handle/321008/173841]  
专题大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者Zhang, Sufang; Zhao, Zongbao K.
作者单位1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
2.Chinese Acad Sci, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China
推荐引用方式
GB/T 7714		 Jiao, Xiang,Zhang, Yue,Liu, Xiangjian,et al. Developing a CRISPR/Cas9 System for Genome Editing in the Basidiomycetous Yeast Rhodosporidium toruloides[J]. BIOTECHNOLOGY JOURNAL,2019,14(7):7.
APA Jiao, Xiang,Zhang, Yue,Liu, Xiangjian,Zhang, Qi,Zhang, Sufang,&Zhao, Zongbao K..(2019).Developing a CRISPR/Cas9 System for Genome Editing in the Basidiomycetous Yeast Rhodosporidium toruloides.BIOTECHNOLOGY JOURNAL,14(7),7.
MLA Jiao, Xiang,et al."Developing a CRISPR/Cas9 System for Genome Editing in the Basidiomycetous Yeast Rhodosporidium toruloides".BIOTECHNOLOGY JOURNAL 14.7(2019):7.

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来源：大连化学物理研究所

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