气候变化领域集成服务门户(CCPortal): Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants

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DOI	10.1126/science.aba8853
	Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants
	Walton R.T.; Christie K.A.; Whittaker M.N.; Kleinstiver B.P.
发表日期	2020
ISSN	0036-8075
起始页码	290
结束页码	296
卷号	368 期号:6488
英文摘要	Manipulation of DNA by CRISPR-Cas enzymes requires the recognition of a protospacer-adjacent motif (PAM), limiting target site recognition to a subset of sequences. To remove this constraint, we engineered variants of Streptococcus pyogenes Cas9 (SpCas9) to eliminate the NGG PAM requirement. We developed a variant named SpG that is capable of targeting an expanded set of NGN PAMs, and we further optimized this enzyme to develop a near-PAMless SpCas9 variant named SpRY (NRN and to a lesser extent NYN PAMs). SpRY nuclease and base-editor variants can target almost all PAMs, exhibiting robust activities on a wide range of sites with NRN PAMs in human cells and lower but substantial activity on those with NYN PAMs. Using SpG and SpRY, we generated previously inaccessible disease-relevant genetic variants, supporting the utility of high-resolution targeting across genome editing applications. © 2020 American Association for the Advancement of Science. All rights reserved.
关键词	adenine adenosine CRISPR associated endonuclease Cas9 cytosine guanosine SpG protein SpRY nuclease thymine unclassified drug Cas9 endonuclease Streptococcus pyogenes bacterium enzyme activity genetic engineering genetic variation genome Article CRISPR-CAS9 system DNA sequence enzyme activity gene editing gene targeting genetic variability human mutagenesis nonhuman priority journal protein engineering protein motif protein structure protospacer adjacent motif Streptococcus pyogenes chemistry CRISPR Cas system enzyme specificity gene editing gene targeting genetic predisposition genetics HEK293 cell line procedures protein domain Streptococcus pyogenes CRISPR-Associated Protein 9 CRISPR-Cas Systems Gene Editing Gene Targeting Genetic Predisposition to Disease HEK293 Cells Humans Mutagenesis Protein Domains Substrate Specificity
语种	英语
来源机构	Science
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/133561
推荐引用方式 GB/T 7714	Walton R.T.,Christie K.A.,Whittaker M.N.,et al. Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants[J]. Science,2020,368(6488).
APA	Walton R.T.,Christie K.A.,Whittaker M.N.,&Kleinstiver B.P..(2020).Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants.,368(6488).
MLA	Walton R.T.,et al."Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants".368.6488(2020).