title: Measuring genetic interactions in human cells by RNAi and imaging.
abstract: 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.
authors.affiliation: 1] German Cancer Research Center (DKFZ), Division of Signaling and Functional Genomics, Heidelberg, Germany. [2] Department of Cell and Molecular Biology, Heidelberg University, Heidelberg, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Laufer
authors.affiliation: European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Heidelberg, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Fischer
authors.affiliation: European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Heidelberg, Germany.
authors.forename: Wolfgang
authors.identifier: 0000000204742218
authors.initials: W
authors.lastname: Huber
authors.affiliation: 1] German Cancer Research Center (DKFZ), Division of Signaling and Functional Genomics, Heidelberg, Germany. [2] Department of Cell and Molecular Biology, Heidelberg University, Heidelberg, Germany.
authors.forename: Michael
authors.identifier: 000000029458817X
authors.initials: M
authors.lastname: Boutros
mesh_terms: D000818:Animals; D055785:Gene Knockdown Techniques / Q000295:instrumentation / Q000379:methods*; D045325:HCT116 Cells; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D010641:Phenotype; D011506:Proteins / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering; D014162:Transfection / Q000379:methods; D057188:Workflow
chemical_list: D011506:Proteins; D034741:RNA, Small Interfering