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

Parnas, Oren; Jovanovic, Marko; Eisenhaure, Thomas M.; Herbst, Rebecca H.; Dixit, Atray; Ye, Chun Jimmie; Przybylski, Dariusz; Platt, Randall J.; Tirosh, Itay; Sanjana, Neville E.; Shalem, Ophir; Satija, Rahul; Raychowdhury, Raktima; Mertins, Philipp; Carr, Steven A.; Zhang, Feng; Hacohen, Nir; Regev, Aviv

doi:10.1016/j.cell.2015.06.059

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

Published Date:

Jul 16 2015

Publisher's site:

http://dx.doi.org/10.1016/j.cell.2015.06.059 New Site Icon

Source:

Cell. 162(3):675-686.

[PDF-2.33 MB]

Details:

Personal Authors:

Parnas, Oren ; Jovanovic, Marko ; Eisenhaure, Thomas M. ; Herbst, Rebecca H. ; Dixit, Atray ; ... More ▼

Keywords:

[+]

Pubmed ID:

26189680

Pubmed Central ID:

PMC4522370

Description:

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.

Document Type:

Journal Article

Collection(s):

CDC Public Access

Funding:

5DP1-MH100706/DP/NCCDPHP CDC HHS/United States
5R01-DK097768/DK/NIDDK NIH HHS/United States
DP1 MH100706/MH/NIMH NIH HHS/United States
K99 HG008171/HG/NHGRI NIH HHS/United States
K99-HG008171/HG/NHGRI NIH HHS/United States
P50 HG006193/HG/NHGRI NIH HHS/United States
P50 HG006193/HG/NHGRI NIH HHS/United States
R01 MH110049/MH/NIMH NIH HHS/United States
Howard Hughes Medical Institute/United States
Howard Hughes Medical Institute/United States

Supporting Files:

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