Expanding the Potential of CRISPR-Cpf1-Based Genome Editing Technology in the Cyanobacterium Anabaena PCC 7120
文献类型：期刊论文
| 作者 | Zhang, Cheng-Cai2; Zhang, Ju-Yuan2; Xing, Wei-Yue1,2; Wang, Zi-Qian1,2; Xie, Li-Rui2; Lin, Gui-Ming2; Niu, Tian-Cai2 |
| 刊名 | ACS SYNTHETIC BIOLOGY
 |
| 出版日期 | 2019 |
| 卷号 | 8期号:1页码:170-180 |
| 关键词 | CRISPR Cpf1 cyanobacteria genome editing conditional mutant large DNA fragment deletion |
| ISSN号 | 2161-5063 |
| DOI | 10.1021/acssynbio.8b00437 |
| 英文摘要 | CRISPR systems, such as CRISPR-Cas9 and CRISPR-Cpf1, have been successfully used for genome editing in a variety of organisms. Although the technique of CRISPR-Cpf1 has been applied in cyanobacteria recently, its use was limited without exploiting the full potential of such a powerful genetic system. Using the cyanobacterium Anabaena PCC 7120 as a model strain, we improved the tools and designed genetic strategies based on CRISPR-Cpf1, which enabled us to realize genetic experiments that have been so far difficult to do in cyanobacteria. The development includes: (1) a "two-spacers" strategy for single genomic modification, with a success rate close to 100%; (2) rapid multiple genome editing using editing plasmids with different resistance markers; (3) using sacB, a counter-selection marker conferring sucrose sensitivity, to enable the active loss of the editing plasmids and facilitate multiple rounds of genetic modification or phenotypic analysis; (4) manipulation of essential genes by the creation of conditional mutants, using as example, polA encoding the DNA polymerase I essential for DNA replication and repair; (5) large DNA fragment deletion, up to 118 kb, from the Anabaena chromosome, corresponding to the largest bacterial chromosomal region removed with CRISPR systems so far. The genome editing vectors and the strategies developed here will expand our ability to study and engineer cyanobacteria, which are extensively used for fundamental studies, biotechnological applications including biofuel production, and synthetic biology research. The vectors developed here have a broad host range, and could be readily used for genetic modification in other microorganisms. |
| 资助项目 | Chinese Academy of Sciences[Y85Z061601] ; Chinese Academy of Sciences[Y65Z021501] ; Key Research Program of Frontier Sciences of the Chinese Academy of Sciences[QYZDJ-SSW-SMC016] ; Recruitment Program of Global Experts of China[Y523011] |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000456633300017 |
| 出版者 | AMER CHEMICAL SOC |
| 源URL | [http://202.127.146.157/handle/2RYDP1HH/6517]  |
| 专题 | 中国科学院武汉植物园 |
| 通讯作者 | Zhang, Ju-Yuan |
| 作者单位 | 1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Chinese Acad Sci, Inst Hydrobiol, Key Lab Algal Biol, Wuhan, Hubei, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Cheng-Cai,Zhang, Ju-Yuan,Xing, Wei-Yue,et al. Expanding the Potential of CRISPR-Cpf1-Based Genome Editing Technology in the Cyanobacterium Anabaena PCC 7120[J]. ACS SYNTHETIC BIOLOGY,2019,8(1):170-180. |
| APA | Zhang, Cheng-Cai.,Zhang, Ju-Yuan.,Xing, Wei-Yue.,Wang, Zi-Qian.,Xie, Li-Rui.,...&Niu, Tian-Cai.(2019).Expanding the Potential of CRISPR-Cpf1-Based Genome Editing Technology in the Cyanobacterium Anabaena PCC 7120.ACS SYNTHETIC BIOLOGY,8(1),170-180. |
| MLA | Zhang, Cheng-Cai,et al."Expanding the Potential of CRISPR-Cpf1-Based Genome Editing Technology in the Cyanobacterium Anabaena PCC 7120".ACS SYNTHETIC BIOLOGY 8.1(2019):170-180. |
入库方式： OAI收割
来源：武汉植物园
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