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Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting
Guo, Lijun1; Xu, Kun1; Liu, Zhiyuan1; Zhang, Cunfang1,2; Xin, Ying1; Zhang, Zhiying1
2015-06-01
Source PublicationANALYTICAL BIOCHEMISTRY
Volume478Pages:131-133
SubtypeArticle
AbstractIn addition to the advantages of scalable, affordable, and easy to engineer, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) technology is superior for multiplex targeting, which is laborious and inconvenient when achieved by cloning multiple gRNA expressing cassettes. Here, we report a simple CRISPR array assembling method which will facilitate multiplex targeting usage. First, the Streptococcus thermophilus CRISPR3/Cas locus was cloned. Second, different CRISPR arrays were assembled with different crRNA spacers. Transformation assays using different Escherichia coli strains demonstrated efficient plasmid DNA targeting, and we achieved targeting efficiency up to 95% with an assembled CRISPR array with three crRNA spacers. (C) 2015 Elsevier Inc. All rights reserved.; In addition to the advantages of scalable, affordable, and easy to engineer, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) technology is superior for multiplex targeting, which is laborious and inconvenient when achieved by cloning multiple gRNA expressing cassettes. Here, we report a simple CRISPR array assembling method which will facilitate multiplex targeting usage. First, the Streptococcus thermophilus CRISPR3/Cas locus was cloned. Second, different CRISPR arrays were assembled with different crRNA spacers. Transformation assays using different Escherichia coli strains demonstrated efficient plasmid DNA targeting, and we achieved targeting efficiency up to 95% with an assembled CRISPR array with three crRNA spacers. (C) 2015 Elsevier Inc. All rights reserved.
KeywordCas9 Crispr Array Crispr/cas Multiplex Targeting Streptococcus Thermophilus
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS KeywordCAS SYSTEMS ; VECTOR ; IMMUNITY
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Analytical
WOS IDWOS:000353839900020
Citation statistics
Cited Times:7[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://210.75.249.4/handle/363003/5551
Collection中国科学院西北高原生物研究所
Affiliation1.Northwest A&F Univ, Coll Anim Sci & Technol, Yangling 712100, Shaanxi, Peoples R China
2.Chinese Acad Sci, Northwest Inst Plateau Biol, Key Lab Adaptat & Evolut Plateau Biota, Xining 810008, Qinghai, Peoples R China
Recommended Citation
GB/T 7714
Guo, Lijun,Xu, Kun,Liu, Zhiyuan,et al. Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting[J]. ANALYTICAL BIOCHEMISTRY,2015,478:131-133.
APA Guo, Lijun,Xu, Kun,Liu, Zhiyuan,Zhang, Cunfang,Xin, Ying,&Zhang, Zhiying.(2015).Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting.ANALYTICAL BIOCHEMISTRY,478,131-133.
MLA Guo, Lijun,et al."Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting".ANALYTICAL BIOCHEMISTRY 478(2015):131-133.
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