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 (US) |
|---|---|
| Pages (from-to) | 675-686 |
| Number of pages | 12 |
| Journal | Cell |
| Volume | 162 |
| Issue number | 3 |
| DOIs | |
| State | Published - Aug 1 2015 |
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ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
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.
In: Cell, Vol. 162, No. 3, 01.08.2015, p. 675-686.Research output: Contribution to journal › Article
}
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 - Article
VL - 162
SP - 675
EP - 686
JO - Cell
JF - Cell
SN - 0092-8674
IS - 3
ER -