A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks
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Regev_A Genome-Wide Crispr.pdf
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Author(s)
Parnas, Oren
Jovanovic, Marko
Eisenhaure, Thomas M.
Herbst, Rebecca H.
Ye, Chun Jimmie
Przybylski, Dariusz
Tirosh, Itay
Satija, Rahul
Raychowdhury, Raktima
Mertins, Philipp
Date Issued
July 2015
Journal
Cell
Publisher
Elsevier
Citation
Parnas, Oren, Marko Jovanovic, Thomas M. Eisenhaure, Rebecca H. Herbst, Atray Dixit, Chun Jimmie Ye, Dariusz Przybylski, et al. “A Genome-Wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks.” Cell 162, no. 3 (July 2015): 675–686.
Version
Author's final manuscript
Abstract
Finding the components of cellular circuits and determining their functions systematically remains a major challenge in mammalian cells. Here, we introduced genome-wide pooled CRISPR-Cas9 libraries into dendritic cells (DCs) to identify genes that control the induction of tumor necrosis factor (Tnf) by bacterial lipopolysaccharide (LPS), a key process in the host response to pathogens, mediated by the Tlr4 pathway. We found many of the known regulators of Tlr4 signaling, as well as dozens of previously unknown candidates that we validated. By measuring protein markers and mRNA profiles in DCs that are deficient in known or candidate genes, we classified the genes into three functional modules with distinct effects on the canonical responses to LPS and highlighted functions for the PAF complex and oligosaccharyltransferase (OST) complex. Our findings uncover new facets of innate immune circuits in primary cells and provide a genetic approach for dissection of mammalian cell circuits.
MIT Department
Institute for Medical Engineering and Science
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
McGovern Institute for Brain Research at MIT
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Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/j.cell.2015.06.059
