Functional genomics platform for pooled screening and generation of mammalian genetic interaction maps.

ABSTRACT: Systematic genetic interaction maps in microorganisms are powerful tools for identifying functional relationships between genes and for defining the function of uncharacterized genes. We have recently implemented this strategy in mammalian cells as a two-stage approach. First, genes of interest are robustly identified in a pooled genome-wide screen using complex shRNA libraries. Second, phenotypes for all pairwise combinations of 'hit' genes are measured in a double-shRNA screen and used to construct a genetic interaction map. Our protocol allows for rapid pooled screening under various conditions without a requirement for robotics, in contrast to arrayed approaches. Each round of screening can be implemented in ∼2 weeks, with additional time for analysis and generation of reagents. We dis

SUBMITTER: Kampmann M 

PROVIDER: S-EPMC4144868 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature