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A Genome-wide CRISPR screen in primary immune cells to dissect regulatory networks

Item Type:Article
Title:A Genome-wide CRISPR screen in primary immune cells to dissect regulatory networks
Creators Name:Parnas, O. and Jovanovic, M. and Eisenhaure, T.M. and Herbst, R.H. and Dixit, A. and Ye, C.J. and Przybylski, D. and Platt, R.J. and Tirosh, I. and Sanjana, N.E. and Shalem, O. and Satija, R. and Raychowdhury, R. and Mertins, P. and Carr, S.A. and Zhang, F. and Hacohen, N. and Regev, A.
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.
Keywords:Bone Marrow Cells, CRISPR-Cas Systems, Cell Differentiation, Cell Survival, Dendritic Cells, Gene Knockout Techniques, Gene Regulatory Networks, Genetic Techniques, Hexosyltransferases, Innate Immunity, Membrane Proteins, Toll-Like Receptor 4, Transgenic Mice, Tumor Necrosis Factor-alpha, Animals, Mice
Publisher:Cell Press
Page Range:675-686
Date:30 July 2015
Official Publication:https://doi.org/10.1016/j.cell.2015.06.059
PubMed:View item in PubMed

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