@article{9dc32c0572774215ba4dc0938f2d6851,
title = "A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks",
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.",
author = "Oren Parnas and Marko Jovanovic and Eisenhaure, {Thomas M.} and Herbst, {Rebecca H.} and Atray Dixit and Ye, {Chun Jimmie} and Dariusz Przybylski and Platt, {Randall J.} and Itay Tirosh and Sanjana, {Neville E.} and Ophir Shalem and Rahul Satija and Raktima Raychowdhury and Philipp Mertins and Carr, {Steven A.} and Feng Zhang and Nir Hacohen and Aviv Regev",
note = "Funding Information: We thank Max Artyomov and Laurie Glimcher for help and data related to Xbp1 targets and Jonathan Weissman for very helpful discussions on ER stress. We thank the Broad{\textquoteright}s Genomic Perturbation Platform for help in design of the secondary library and Broad Technology labs (Tarjei Mikkelssen) for help with synthesis of the secondary library. We thank Dave Gennert, Carl De Boer, Alex Shalek, John Trombetta, Schraga Schwartz, Maxwell Mumbach, Dawn Thompson, Timothy Tickle, Brian Haas, Chloe Villani, and all members of the Regev, Hacohen, Carr, and Zhang groups for input and discussions. We thank Terry Means for discussions and advice. We thank Leslie Gaffney for help with figure preparation and the Broad Genomics Platform for all sequencing. FACS sorting was performed at the Bauer Core Laboratory, Harvard FAS Center for Systems Biology, with great help from Patricia Rogers. This work was supported by NHGRI CEGS P50 HG006193 (A.R., N.H., and S.A.C.) and Broad Institute Funds. A.R. was supported by the Klarman Cell Obervatory and HHMI. N.H. was supported by the MGH Research Scholars Program. M.J. was supported by fellowships of the Swiss National Science Foundation for advanced researchers (SNF) and the Marie Sklodowska-Curie IOF. F.Z. is supported by the National Institutes of Health through NIMH (5DP1-MH100706) and NIDDK (5R01-DK097768), a Waterman Award from the National Science Foundation, the Keck, New York Stem Cell, Damon Runyon, Searle Scholars, Merkin, and Vallee Foundations, and Bob Metcalfe. F.Z. is a New York Stem Cell Foundation Robertson Investigator. N.E.S. is supported by a Simons Center for the Social Brain Postdoctoral Fellowship and NIH NHGRI award K99-HG008171. O.S. is a fellow of the Klarman Cell Observatory. R.J.P. is supported by a National Science Foundation Graduate Research Fellowship under grant number 1122374. I.T. was supported by a Human Frontier Science Program fellowship. A.D. is supported by the National Space Biomedical Research Institute through NASA NCC 9-58 and the National Defense Science and Engineering Fellowship. R.H.H. receives support from the Herchel Smith Fellowship. A.R. is on the scientific advisory board for Thermo Fisher Scientific. F.Z. is a founder and scientific advisor of Editas Medicine and a scientific advisor for Horizon Discovery. Publisher Copyright: {\textcopyright} 2015 Elsevier Inc.",
year = "2015",
month = aug,
day = "1",
doi = "10.1016/j.cell.2015.06.059",
language = "English (US)",
volume = "162",
pages = "675--686",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "3",
}