用于确定非编码基因组功能的高通量方法。
High-Throughput Approaches to Pinpoint Function within the Noncoding Genome.
作者信息
Montalbano Antonino, Canver Matthew C, Sanjana Neville E
机构信息
New York Genome Center, New York, NY, USA; Department of Biology, New York University, New York, NY, USA.
Harvard Medical School, Boston, MA, USA.
出版信息
Mol Cell. 2017 Oct 5;68(1):44-59. doi: 10.1016/j.molcel.2017.09.017.
The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas nuclease system is a powerful tool for genome editing, and its simple programmability has enabled high-throughput genetic and epigenetic studies. These high-throughput approaches offer investigators a toolkit for functional interrogation of not only protein-coding genes but also noncoding DNA. Historically, noncoding DNA has lacked the detailed characterization that has been applied to protein-coding genes in large part because there has not been a robust set of methodologies for perturbing these regions. Although the majority of high-throughput CRISPR screens have focused on the coding genome to date, an increasing number of CRISPR screens targeting noncoding genomic regions continue to emerge. Here, we review high-throughput CRISPR-based approaches to uncover and understand functional elements within the noncoding genome and discuss practical aspects of noncoding library design and screen analysis.
成簇规律间隔短回文重复序列（CRISPR）-Cas核酸酶系统是一种强大的基因组编辑工具，其简单的可编程性推动了高通量遗传和表观遗传研究。这些高通量方法为研究人员提供了一个工具包，不仅可用于对蛋白质编码基因进行功能研究，还可用于对非编码DNA进行功能研究。从历史上看，非编码DNA缺乏对蛋白质编码基因所进行的详细表征，这在很大程度上是因为一直没有一套强大的方法来干扰这些区域。尽管到目前为止，大多数高通量CRISPR筛选都集中在编码基因组上，但越来越多针对非编码基因组区域的CRISPR筛选不断涌现。在这里，我们综述了基于CRISPR的高通量方法，以揭示和理解非编码基因组中的功能元件，并讨论非编码文库设计和筛选分析的实际问题。
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