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CRISPR-Cas9~(D10A) nickase-assisted base editing in solvent producer Clostridium beijerinckii

李琦  Fran?ois M.Seys  Nigel P.Minton  Junjie Yang  Yu Jiang  Weihong Jiang  杨晟  
【摘要】:Clostridium beijerinckii is a potentially important industrial microorganism as it can synthesize valuable chemicals and fuels from various carbon sources. The establishment of convenient to use, effective gene tools with which the organism can be rapidly modified is essential if its full potential is to be realized. Here, we developed a genomic editing tool(pCBEclos) for use in C. beijerinckii based on the fusion of cytidine deaminase(Apobec1), Cas9 D10 A nickase and uracil DNA glycosylase inhibitor(UGI). Apobec1 and UGI are guided to the target site where they introduce specific base-pair substitutions through the conversion of C·G to T·A. By appropriate choice of target sequence, these nucleotide changes are capable of creating missense mutation or null mutations in a gene. Through optimization of pCBEclos, the system derived, pCBEclos-opt, has been used to rapidly generate four different mutants in C. beijerinckii, in pyrE, xylR, spo0 A and araR. The efficiency of the system was such that they could sometimes be directly obtained following transformation, otherwise only requiring one single re-streaking step. Whilst CRISPR-Cas9 nickase systems, such as pNICKclos2.0, have previously been reported in C. beijerinckii, pCBEclos-opt does not rely on homologous recombination, a process that is intrinsically inefficient in clostridia such as C. beijerinckii. As a consequence, bulky editing templates do not need to be included in the knock-out plasmids. This both reduces plasmid size and makes their construction simpler, e.g., whereas the assembly of pNICKclos2.0 requires six primers for the assembly of a typical knock-out plasmid, pCBEclos-opt requires just two primers. The pCBEclos-opt plasmid established here represents a powerful new tool for genome editing in C. beijerinckii, which should be readily applicable to other clostridial species.

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3 Xin LUO;Min LI;Bing SU;;Application of the genome editing tool CRISPR/Cas9 in non-human primates[J];Zoological Research;2016年04期
4 Yongping Zhang;Wei Mu;Haoyi Wang;;Gene editing in T cell therapy[J];Journal of Genetics and Genomics;2017年09期
5 Weng Yongqing;;CHINA EDITOLOGY SOCIETY OF SCIENCE PERIODICALS ESTABLISHED IN BEIJING[J];Journal of Chemical Industry and Engineering;1987年02期
6 Yuyan Yang;Yue Huang;;The CRIPSR/Cas gene-editing system——an immature but useful toolkit for experimental and clinical medicine[J];Animal Models and Experimental Medicine;2019年01期
7 Xiangbing Meng;Xixun Hu;Qing Liu;Xiaoguang Song;Caixia Gao;Jiayang Li;Kejian Wang;;Robust genome editing of CRISPR-Cas9 at NAG PAMs in rice[J];Science China(Life Sciences);2018年01期
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9 Yiyu Chen;Zhiping Wang;Hanwen Ni;Yong Xu;Qijun Chen;Linjian Jiang;;CRISPR/Cas9-mediated base-editing system efficiently generates gain-of-function mutations in Arabidopsis[J];Science China(Life Sciences);2017年05期
10 KWON Ji-Yong;LEE In-Kwon;;Graphical interface for motion editing using procedure visualization[J];Science China(Information Sciences);2011年06期
11 ;Adaptive sampling for mesh spectrum editing[J];Journal of Zhejiang University Science A(Science in Engineering);2006年07期
12 Ming SHAO;Tian-Rui XU;Ce-Shi CHEN;;The big bang of genome editing technology: development and application of the CRISPR/Cas9 system in disease animal models[J];Zoological Research;2016年04期
13 Jingying Li;Jinman Luo;Meilian Xu;Shaoya Li;Jiahui Zhang;Huiyuan Li;Lei Yan;Yunde Zhao;Lanqin Xia;;Plant genome editing using xCas9 with expanded PAM compatibility[J];Journal of Genetics and Genomics;2019年05期
14 Jianguo Zhao;Liangxue Lai;Weizhi Ji;Qi Zhou;;Genome editing in large animals: current status and future prospects[J];National Science Review;2019年03期
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