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Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting
Guo, Lijun ; Xu, Kun ; Liu, Zhiyuan ; Zhang, Cunfang ; Xin, Ying ; Zhang, Zhiying
2015-06-01
发表期刊ANALYTICAL BIOCHEMISTRY
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.
文献类型期刊论文
条目标识符http://210.75.249.4/handle/363003/22429
专题中国科学院西北高原生物研究所
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.
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.
MLA Guo, Lijun,et al."Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting".ANALYTICAL BIOCHEMISTRY (2015).
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