全基因组规模的CRISPR混合筛选
Genome-scale CRISPR pooled screens.
作者信息
Sanjana Neville E
机构信息
New York Genome Center, New York, NY 10013, USA; Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10012, USA.
出版信息
Anal Biochem. 2017 Sep 1;532:95-99. doi: 10.1016/j.ab.2016.05.014. Epub 2016 Jun 1.
Abstract
Genome editing technologies such as clustered regularly interspaced short palindromic repeats (CRISPR) systems have ushered in a new era of targeted DNA manipulation. The easy programmability of CRISPR using short oligonucleotides enables rapid synthesis of large-scale libraries for functional genetic screens. Here we present fundamental concepts and methods for pooled CRISPR screens and review biological results from recent genome-scale loss-of-function and gain-of-function screens. We also discuss new frontiers in pooled screens, including novel effector domains for functional screens and applications in the noncoding genome.
摘要
诸如成簇规律间隔短回文重复序列（CRISPR）系统等基因组编辑技术开创了靶向DNA操纵的新时代。CRISPR利用短寡核苷酸实现的易于编程性，使得能够快速合成用于功能基因筛选的大规模文库。在此，我们介绍了汇集式CRISPR筛选的基本概念和方法，并回顾了近期全基因组规模功能丧失和功能获得筛选的生物学结果。我们还讨论了汇集式筛选的新前沿，包括用于功能筛选的新型效应结构域以及在非编码基因组中的应用。
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