Measuring genetic interactions in human cells by RNAi and imaging

被引：15
|
作者：
Laufer, Christina [1 ,2 ]
Fischer, Bernd [3 ]
Huber, Wolfgang [3 ]
Boutros, Michael [1 ,2 ]
机构：
[1] German Canc Res Ctr, Div Signaling & Funct Genom, Heidelberg, Germany
[2] Heidelberg Univ, Dept Cell & Mol Biol, Heidelberg, Germany
[3] European Mol Biol Lab, Genome Biol Unit, D-69012 Heidelberg, Germany
来源：
NATURE PROTOCOLS | 2014年 / 9卷 / 10期
基金：
欧洲研究理事会;
关键词：
NETWORKS; PHENOTYPES;
D O I：
10.1038/nprot.2014.160
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Observation of how genetic interactions modulate phenotypes is a powerful method for dissecting their underlying molecular and functional networks. Whereas in model organisms genetic interaction studies are well established, systematic analysis of genetic interactions in human cells has remained challenging. Here we provide a detailed protocol for large-scale mapping of genetic interactions in human cells using a high-throughput phenotyping approach. Pairwise gene product depletion is induced by siRNA-mediated knockdown, and the resulting phenotypes are quantified by automated imaging and computational analysis to provide the basis for detecting genetic interactions between all pairs of genes tested. The whole workflow, depending on the size of the experiment, takes 3 or more weeks to complete.
引用
收藏
页码：2341 / 2353
页数：13
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