Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting

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	Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting
	Guo, Lijun 1; Xu, Kun 1; Liu, Zhiyuan 1; Zhang, Cunfang 1,2; Xin, Ying 1; Zhang, Zhiying 1
	2015-06-01
发表期刊	ANALYTICAL BIOCHEMISTRY
卷号	478 页码:131-133
文章类型	Article
摘要	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.; 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.
关键词	Cas9 Crispr Array Crispr/cas Multiplex Targeting Streptococcus Thermophilus
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
关键词[WOS]	CAS SYSTEMS ; VECTOR ; IMMUNITY
收录类别	SCI
语种	英语
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
WOS类目	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Analytical
WOS记录号	WOS:000353839900020
引用统计	被引频次：7[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://210.75.249.4/handle/363003/5551
专题	中国科学院西北高原生物研究所
作者单位	1.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
推荐引用方式 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.