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Expanding the Potential of CRISPR-Cpf1-Based Genome Editing Technology in the Cyanobacterium Anabaena PCC 7120
Niu, Tian-Cai1; Lin, Gui-Ming1; Xie, Li-Rui1; Wang, Zi-Qian1,2; Xing, Wei-Yue1,2; Zhang, Ju-Yuan1; Zhang, Cheng-Cai1
Corresponding AuthorZhang, Ju-Yuan(zhangjuyuan@ihb.ac.cn)
2019
Source PublicationACS SYNTHETIC BIOLOGY
ISSN2161-5063
Volume8Issue:1Pages:170-180
AbstractCRISPR 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.
KeywordCRISPR Cpf1 cyanobacteria genome editing conditional mutant large DNA fragment deletion
DOI10.1021/acssynbio.8b00437
Funding OrganizationChinese Academy of Sciences ; Key Research Program of Frontier Sciences of the Chinese Academy of Sciences ; Recruitment Program of Global Experts of China ; Chinese Academy of Sciences ; Key Research Program of Frontier Sciences of the Chinese Academy of Sciences ; Recruitment Program of Global Experts of China
Indexed BySCI ; SCI
Language英语
Funding ProjectChinese 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 Research AreaBiochemistry & Molecular Biology
WOS SubjectBiochemical Research Methods
WOS IDWOS:000456633300017
WOS KeywordDNA-POLYMERASE-I ; SP-STRAIN PCC-7120 ; HETEROCYST PATTERN ; GENE ; CRISPR-CAS9 ; EXPRESSION ; CPF1 ; TOOL ; ENDONUCLEASE ; PURIFICATION
PublisherAMER CHEMICAL SOC
Funding OrganizationChinese Academy of Sciences ; Key Research Program of Frontier Sciences of the Chinese Academy of Sciences ; Recruitment Program of Global Experts of China ; Chinese Academy of Sciences ; Key Research Program of Frontier Sciences of the Chinese Academy of Sciences ; Recruitment Program of Global Experts of China
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/28766
Corresponding AuthorZhang, Ju-Yuan
Affiliation1.Chinese Acad Sci, Inst Hydrobiol, Key Lab Algal Biol, Wuhan, Hubei, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Niu, Tian-Cai,Lin, Gui-Ming,Xie, Li-Rui,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 Niu, Tian-Cai.,Lin, Gui-Ming.,Xie, Li-Rui.,Wang, Zi-Qian.,Xing, Wei-Yue.,...&Zhang, Cheng-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 Niu, Tian-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.
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