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

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

doi:10.1016/j.cell.2015.06.059

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

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

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	675-686
Number of pages	12
Journal	Cell
Volume	162
Issue number	3
DOIs	https://doi.org/10.1016/j.cell.2015.06.059
State	Published - 1 Aug 2015
Externally published	Yes

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This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

All Science Journal Classification (ASJC) codes

General Biochemistry,Genetics and Molecular Biology

Access to Document

10.1016/j.cell.2015.06.059

Cite this

Parnas, O., Jovanovic, M., Eisenhaure, T. M., Herbst, R. H., Dixit, A., Ye, C. J., Przybylski, D., Platt, R. J., Tirosh, I., Sanjana, N. E., Shalem, O., Satija, R., Raychowdhury, R., Mertins, P., Carr, S. A., Zhang, F., Hacohen, N., & Regev, A. (2015). A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks. Cell, 162(3), 675-686. https://doi.org/10.1016/j.cell.2015.06.059

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title = "A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks",

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.",

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

note = "Publisher Copyright: {\textcopyright} 2015 Elsevier Inc.",

year = "2015",

month = aug,

day = "1",

doi = "10.1016/j.cell.2015.06.059",

language = "الإنجليزيّة",

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pages = "675--686",

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Parnas, O, Jovanovic, M, Eisenhaure, TM, Herbst, RH, Dixit, A, Ye, CJ, Przybylski, D, Platt, RJ, Tirosh, I, Sanjana, NE, Shalem, O, Satija, R, Raychowdhury, R, Mertins, P, Carr, SA, Zhang, F, Hacohen, N & Regev, A 2015, 'A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks', Cell, vol. 162, no. 3, pp. 675-686. https://doi.org/10.1016/j.cell.2015.06.059

TY - JOUR

T1 - A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks

AU - Parnas, Oren

AU - Jovanovic, Marko

AU - Eisenhaure, Thomas M.

AU - Herbst, Rebecca H.

AU - Dixit, Atray

AU - Ye, Chun Jimmie

AU - Przybylski, Dariusz

AU - Platt, Randall J.

AU - Tirosh, Itay

AU - Sanjana, Neville E.

AU - Shalem, Ophir

AU - Satija, Rahul

AU - Raychowdhury, Raktima

AU - Mertins, Philipp

AU - Carr, Steven A.

AU - Zhang, Feng

AU - Hacohen, Nir

AU - Regev, Aviv

PY - 2015/8/1

Y1 - 2015/8/1

N2 - 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.

AB - 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.

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U2 - 10.1016/j.cell.2015.06.059

DO - 10.1016/j.cell.2015.06.059

M3 - مقالة

C2 - 26189680

SN - 0092-8674

VL - 162

SP - 675

EP - 686

JO - Cell

JF - Cell

IS - 3

ER -

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

Abstract

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