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

Source:

Cell. 162(3):675-686.

[PDF-2.33 MB]

Details:

Alternative Title:

Cell

Personal Author:

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

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.

Subject:

[+]

Pubmed ID:

26189680

Pubmed Central ID:

PMC4522370

Document Type:

Journal Article

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 ; ... More ▼

Collection(s):

CDC Public Access

Main Document Checksum:

[+]

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