在细胞中使用RNAi与基于CRISPR的基因敲除相结合的合成致死筛选
Synthetic Lethality Screens Using RNAi in Combination with CRISPR-based Knockout in Cells.
作者信息
Housden Benjamin E, Nicholson Hilary E, Perrimon Norbert
机构信息
Harvard Medical School, Boston, USA.
Dana-Farber Cancer Institute, Boston, USA.
出版信息
Bio Protoc. 2017 Feb 5;7(3). doi: 10.21769/BioProtoc.2119.
Abstract
A synthetic lethal interaction is a type of genetic interaction where the disruption of either of two genes individually has little effect but their combined disruption is lethal. Knowledge of synthetic lethal interactions can allow for elucidation of network structure and identification of candidate drug targets for human diseases such as cancer. In , combinatorial gene disruption has been achieved previously by combining multiple RNAi reagents. Here we describe a protocol for high-throughput combinatorial gene disruption by combining CRISPR and RNAi. This approach previously resulted in the identification of highly reproducible and conserved synthetic lethal interactions (Housden , 2015).
摘要
合成致死相互作用是一种基因相互作用类型，其中两个基因单独被破坏时影响很小，但它们的联合破坏却是致命的。了解合成致死相互作用有助于阐明网络结构，并识别癌症等人类疾病的候选药物靶点。在以往的研究中，通过组合多种RNAi试剂实现了组合基因破坏。在此，我们描述了一种通过结合CRISPR和RNAi进行高通量组合基因破坏的方案。该方法此前已导致识别出高度可重复和保守的合成致死相互作用（豪斯登，2015年）。
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