Journal
CELL
Volume 179, Issue 3, Pages 787-+Publisher
CELL PRESS
DOI: 10.1016/j.cell.2019.09.016
Keywords
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Funding
- Simons Center Seed Grant from MIT
- Broad Institute
- BN10 program
- National Human Genome Research Institute [HG009283, HG006193]
- Burroughs Wellcome Fund
- NIH [1R01-HG009761, 1R01-MH110049, 1DP1-HL141201]
- Howard Hughes Medical Institute
- New York Stem Cell Foundation
- Simons Foundation
- Paul G. Allen Family Foundation
- Vallee Foundation
- Poitras Center for Affective Disorders Research at MIT
- Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT
- Swedish Research Council [2015-06403]
- Fannie and John Hertz Foundation Fellowship
- NSF Graduate Research Fellowship
- EMBO Long-Term Fellowship [ALTF 199-2017]
- Swedish Research Council [2015-06403] Funding Source: Swedish Research Council
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Genetic screens are critical for the systematic identification of genes underlying cellular phenotypes. Pooling gene perturbations greatly improves scalability but is not compatible with imaging of complex and dynamic cellular phenotypes. Here, we introduce a pooled approach for optical genetic screens in mammalian cells. We use targeted in situ sequencing to demultiplex a library of genetic perturbations following image-based phenotyping. We screened a set of 952 genes across millions of cells for involvement in nuclear factor kB (NF-kappa B) signaling by imaging the translocation of RelA (p65) to the nucleus. Screening at a single time point across 3 cell lines recovered 15 known pathway components, while repeating the screen with live-cell imaging revealed a role for Mediator complex subunits in regulating the duration of p65 nuclear retention. These results establish a highly multiplexed approach to imagebased screens of spatially and temporally defined phenotypes with pooled libraries.
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