Crispr-based genome editing and expression control systems in clostridium acetobutylicum and clostridium beijerinckii
文献类型：期刊论文
| 作者 | Li, Qi1,2; Chen, Jun1; Minton, Nigel P.3; Zhang, Ying3; Wen, Zhiqiang1; Liu, Jinle1,2; Yang, Haifeng1,2; Zeng, Zhe1,2; Ren, Xiaodan1,2; Yang, Junjie1 |
| 刊名 | Biotechnology journal
 |
| 出版日期 | 2016-07-01 |
| 卷号 | 11期号:7页码:961-972 |
| 关键词 | Clostridium Crispr-cas9 Gene expression Genome editing Nickase |
| ISSN号 | 1860-6768 |
| DOI | 10.1002/biot.201600053 |
| 通讯作者 | Yang, sheng(syang@sibs.ac.cn) |
| 英文摘要 | Solventogenic clostridia are important industrial microorganisms that produce various chemicals and fuels. effective genetic tools would facilitate physiological studies aimed both at improving our understanding of metabolism and optimizing solvent productivity through metabolic engineering. here we have developed an all-in-one, crispr-based genome editing plasmid, pnickclos, that can be used to achieve successive rounds of gene editing in clostridium acetobutylicum atcc 824 and clostridium beijerinckii ncimb 8052 with efficiencies varying from 6.7% to 100% and 18.8% to 100%, respectively. the plasmid specifies the requisite target-specific guide rna, the gene encoding the streptococcus pyogenes cas9 nickase and the genome editing template encompassing the gene-specific homology arms. it can be used to create single target mutants within three days, with a further two days required for the curing of the pnickclos plasmid ready for a second round of mutagenesis. a s. pyogenes dcas9-mediated gene regulation control system, pdcasclos, was also developed and used in a crispri strategy to successfully repress the expression of spo0a in c. acetobutylicum and c. beijerinckii. the combined application of the established high efficiency crispr-cas9 based genome editing and regulation control systems will greatly accelerate future progress in the understanding and manipulation of metabolism in solventogenic clostridia. |
| WOS关键词 | FERMENTATIVE BUTANOL PRODUCTION ; ESCHERICHIA-COLI ; ATCC 824 ; HOMOLOGOUS RECOMBINATION ; ALLELIC EXCHANGE ; GENE-EXPRESSION ; ACETONE FORMATION ; CAS SYSTEMS ; RNA ; ACTIVATION |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| WOS类目 | Biochemical Research Methods ; Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000379786700011 |
| 出版者 | WILEY-V C H VERLAG GMBH |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2374631 |
| 专题 | 中国科学院大学 |
| 通讯作者 | Yang, Sheng |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, Key Lab Synthet Biol, Shanghai, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China 3.Univ Nottingham, Sch Life Sci, BBSRC EPSRC Synthet Biol Res Ctr SBRC, Clostridia Res Grp, Nottingham NG7 2RD, England 4.Shanghai Res & Dev Ctr Ind Biotechnol, Shanghai, Peoples R China 5.Jiangsu Natl Synerget Innovat Ctr Adv Mat, Nanjing, Jiangsu, Peoples R China |
| 推荐引用方式 GB/T 7714 | Li, Qi,Chen, Jun,Minton, Nigel P.,et al. Crispr-based genome editing and expression control systems in clostridium acetobutylicum and clostridium beijerinckii[J]. Biotechnology journal,2016,11(7):961-972. |
| APA | Li, Qi.,Chen, Jun.,Minton, Nigel P..,Zhang, Ying.,Wen, Zhiqiang.,...&Yang, Sheng.(2016).Crispr-based genome editing and expression control systems in clostridium acetobutylicum and clostridium beijerinckii.Biotechnology journal,11(7),961-972. |
| MLA | Li, Qi,et al."Crispr-based genome editing and expression control systems in clostridium acetobutylicum and clostridium beijerinckii".Biotechnology journal 11.7(2016):961-972. |
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来源：中国科学院大学
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