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

被引：342

作者：

Parnas, Oren ^{[1
]}

Jovanovic, Marko ^{[1
]}

Eisenhaure, Thomas M. ^{[1
,2
]}

Herbst, Rebecca H. ^{[1
,3
]}

Dixit, Atray ^{[1
,4
]}

Ye, Chun Jimmie ^{[5
]}

Przybylski, Dariusz ^{[1
]}

Platt, Randall J. ^{[1
,6
,7
,8
]}

Tirosh, Itay ^{[1
]}

Sanjana, Neville E. ^{[1
,6
,8
,9
]}

Shalem, Ophir ^{[1
,6
]}

Satija, Rahul ^{[10
,11
]}

Raychowdhury, Raktima ^{[1
]}

Mertins, Philipp ^{[1
]}

Carr, Steven A. ^{[1
]}

Zhang, Feng ^{[1
,6
,7
,8
,9
]}

Hacohen, Nir ^{[1
,2
,12
]}

Regev, Aviv ^{[1
,13
]}

机构：

[1] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[2] Massachusetts Gen Hosp, Ctr Immunol & Inflammatory Dis, Charlestown, MA 02129 USA

[3] Harvard Univ, Sch Med, Dept Syst Biol, Boston, MA 02114 USA

[4] Harvard MIT Div Hlth Sci & Technol, Cambridge, MA 02139 USA

[5] Univ Calif San Francisco, Dept Bioengn & Therapeut Sci, Dept Epidemiol & Biostat, Inst Human Genet, San Francisco, CA 94143 USA

[6] MIT, McGovern Inst Brain Res, Cambridge, MA 02139 USA

[7] MIT, Dept Brain & Cognit Sci, Cambridge, MA 02139 USA

[8] MIT, Dept Biol Engn, Cambridge, MA 02139 USA

[9] Broad Inst MIT & Harvard, Stanley Ctr Psychiat Res, Cambridge, MA 02142 USA

[10] New York Genome Ctr, New York, NY 10013 USA

[11] NYU, Ctr Genom & Syst Biol, New York, NY 10012 USA

[12] Harvard Univ, Sch Med, Dept Med, Boston, MA 02114 USA

[13] MIT, Howard Hughes Med Inst, Dept Biol, Cambridge, MA 02140 USA

来源：

CELL | 2015年 / 162卷 / 03期

基金：

美国国家卫生研究院; 美国国家科学基金会; 瑞士国家科学基金会;

关键词：

GENE-EXPRESSION; PROTEIN; DIFFERENTIATION; CANCER; ACTIVATION; RESPONSES; ALPHA;

D O I：

10.1016/j.cell.2015.06.059

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：675 / 686

页数：12

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