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

Details

• Alternative Title:
  Cell
• Personal Author:
  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
• 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.
• Subjects:
  Animals Article Bone Marrow Cells Cell Differentiation Cell Survival CRISPR-Cas Systems Dendritic Cells Gene Knockout Techniques Gene Regulatory Networks Genetic Techniques Hexosyltransferases Immunity, Innate Membrane Proteins Mice Mice, Transgenic Toll-Like Receptor 4 Tumor Necrosis Factor-alpha

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• Source:
  Cell. 162(3):675-686.
• 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 ; P50 HG006193/HG/NHGRI NIH HHS/United States ; R01 MH110049/MH/NIMH NIH HHS/United States ; Howard Hughes Medical Institute/United States
• Volume:
  162
• Issue:
  3
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:cec8821c0d512f8902acf582caa822e29086aab74885b3e87a5427212eee778c
• Download URL:
  https://stacks.cdc.gov/view/cdc/34824/cdc_34824_DS1.pdf
• File Type:
  [PDF - 2.33 MB ]