气候变化领域集成服务门户(CCPortal): A versatile platform for locus-scale genome rewriting and verification

CCPortal

DOI	10.1073/pnas.2023952118
	A versatile platform for locus-scale genome rewriting and verification
	Brosh R.; Laurent J.M.; Ordoñez R.; Huang E.; Hogan M.S.; Hitchcock A.M.; Mitchell L.A.; Pinglay S.; Cadley J.A.; Luther R.D.; Truong D.M.; Boeke J.D.; Maurano M.T.
发表日期	2021
ISSN	00278424
卷号	118 期号:10
英文摘要	Routine rewriting of loci associated with human traits and diseases would facilitate their functional analysis. However, existing DNA integration approaches are limited in terms of scalability and portability across genomic loci and cellular contexts. We describe Big-IN, a versatile platform for targeted integration of large DNAs into mammalian cells. CRISPR/Cas9-mediated targeting of a landing pad enables subsequent recombinase-mediated delivery of variant payloads and efficient positive/negative selection for correct clones in mammalian stem cells. We demonstrate integration of constructs up to 143 kb, and an approach for one-step scarless delivery. We developed a staged pipeline combining PCR genotyping and targeted capture sequencing for economical and comprehensive verification of engineered stem cells. Our approach should enable combinatorial interrogation of genomic functional elements and systematic locus-scale analysis of genome function. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Genome engineering; Genome writing; Regulatory genomics; Stem cells; Synthetic biology
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180417
作者单位	Institute for Systems Genetics, NYU Langone Health, New York, NY 10016, United States; Department of Biochemistry and Molecular Pharmacology, NYU Langone Health, New York, NY 10016, United States; Department of Biomedical Engineering, NYU Tandon School of Engineering, Brooklyn, NY 11201, United States; Department of Pathology, NYU Langone Health, New York, NY 10016, United States
推荐引用方式 GB/T 7714	Brosh R.,Laurent J.M.,Ordoñez R.,et al. A versatile platform for locus-scale genome rewriting and verification[J],2021,118(10).
APA	Brosh R..,Laurent J.M..,Ordoñez R..,Huang E..,Hogan M.S..,...&Maurano M.T..(2021).A versatile platform for locus-scale genome rewriting and verification.Proceedings of the National Academy of Sciences of the United States of America,118(10).
MLA	Brosh R.,et al."A versatile platform for locus-scale genome rewriting and verification".Proceedings of the National Academy of Sciences of the United States of America 118.10(2021).