A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks.

Cell
Authors
Keywords
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.

Year of Publication
2015
Journal
Cell
Volume
162
Issue
3
Pages
675-86
Date Published
2015 Jul 30
ISSN
1097-4172
URL
DOI
10.1016/j.cell.2015.06.059
PubMed ID
26189680
PubMed Central ID
PMC4522370
Links
Grant list
R01 MH110049 / MH / NIMH NIH HHS / United States
P30 DK043351 / DK / NIDDK NIH HHS / United States
K99 HG008171 / HG / NHGRI NIH HHS / United States
5DP1-MH100706 / DP / NCCDPHP CDC HHS / United States
K99-HG008171 / HG / NHGRI NIH HHS / United States
Howard Hughes Medical Institute / United States
DP1 MH100706 / MH / NIMH NIH HHS / United States
P50 HG006193 / HG / NHGRI NIH HHS / United States
R01 DK097768 / DK / NIDDK NIH HHS / United States
5R01-DK097768 / DK / NIDDK NIH HHS / United States