PMID- 33904395 OWN - NLM STAT- MEDLINE DCOM- 20211027 LR - 20240402 IS - 2050-084X (Electronic) IS - 2050-084X (Linking) VI - 10 DP - 2021 Apr 27 TI - CRISPR-based functional genomics in human dendritic cells. LID - 10.7554/eLife.65856 [doi] LID - e65856 AB - Dendritic cells (DCs) regulate processes ranging from antitumor and antiviral immunity to host-microbe communication at mucosal surfaces. It remains difficult, however, to genetically manipulate human DCs, limiting our ability to probe how DCs elicit specific immune responses. Here, we develop a CRISPR-Cas9 genome editing method for human monocyte-derived DCs (moDCs) that mediates knockouts with a median efficiency of >94% across >300 genes. Using this method, we perform genetic screens in moDCs, identifying mechanisms by which DCs tune responses to lipopolysaccharides from the human microbiome. In addition, we reveal donor-specific responses to lipopolysaccharides, underscoring the importance of assessing immune phenotypes in donor-derived cells, and identify candidate genes that control this specificity, highlighting the potential of our method to pinpoint determinants of inter-individual variation in immunity. Our work sets the stage for a systematic dissection of the immune signaling at the host-microbiome interface and for targeted engineering of DCs for neoantigen vaccination. CI - (c) 2021, Jost et al. FAU - Jost, Marco AU - Jost M AUID- ORCID: 0000-0002-1369-4908 AD - Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States. AD - Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States. AD - California Institute for Quantitative Biosciences, University of California, San Francisco, San Francisco, United States. AD - Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, United States. FAU - Jacobson, Amy N AU - Jacobson AN AD - Department of Bioengineering, Stanford University, Stanford, United States. AD - ChEM-H, Stanford University, Stanford, United States. FAU - Hussmann, Jeffrey A AU - Hussmann JA AD - Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States. AD - Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States. AD - California Institute for Quantitative Biosciences, University of California, San Francisco, San Francisco, United States. AD - Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, United States. AD - Whitehead Institute for Biomedical Research, Cambridge, United States. FAU - Cirolia, Giana AU - Cirolia G AD - Chan Zuckerberg Biohub, San Francisco, United States. FAU - Fischbach, Michael A AU - Fischbach MA AD - Department of Bioengineering, Stanford University, Stanford, United States. AD - ChEM-H, Stanford University, Stanford, United States. AD - Chan Zuckerberg Biohub, San Francisco, United States. FAU - Weissman, Jonathan S AU - Weissman JS AUID- ORCID: 0000-0003-2445-670X AD - Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States. AD - Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States. AD - California Institute for Quantitative Biosciences, University of California, San Francisco, San Francisco, United States. AD - Whitehead Institute for Biomedical Research, Cambridge, United States. AD - Department of Biology, Massachusetts Institute of Technology, Cambridge, United States. LA - eng SI - GEO/GSE161401 SI - GEO/GSE161466 GR - U01 CA217882/CA/NCI NIH HHS/United States GR - U54 CA224081/CA/NCI NIH HHS/United States GR - HHMI/Howard Hughes Medical Institute/United States GR - DP1 DK113598/DK/NIDDK NIH HHS/United States GR - K99 GM130964/GM/NIGMS NIH HHS/United States GR - R01 DK110174/DK/NIDDK NIH HHS/United States PT - Comparative Study PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20210427 PL - England TA - Elife JT - eLife JID - 101579614 RN - 0 (Cytokines) RN - 0 (Lipopolysaccharides) RN - 0 (TLR4 protein, human) RN - 0 (Toll-Like Receptor 4) RN - EC 3.1.- (CRISPR-Associated Protein 9) SB - IM MH - Bacteroides thetaiotaomicron/immunology MH - CRISPR-Associated Protein 9/*genetics/metabolism MH - *CRISPR-Cas Systems MH - Cells, Cultured MH - *Clustered Regularly Interspaced Short Palindromic Repeats MH - Cytokines/genetics/metabolism MH - Dendritic Cells/drug effects/*immunology/metabolism MH - *Gene Editing MH - Gene Expression Regulation MH - *Genomics MH - Humans MH - Immunity, Innate/drug effects/*genetics MH - Lipopolysaccharides/pharmacology MH - Phenotype MH - Signal Transduction MH - Toll-Like Receptor 4/agonists/genetics/metabolism PMC - PMC8104964 OTO - NOTNLM OT - CRISPR OT - bacteroides thetaiotaomicron OT - dendritic cells OT - functional genomics OT - human OT - immunology OT - infectious disease OT - inflammation OT - inter-individual variation OT - microbiology COIS- MJ consults for Maze Therapeutics. AJ, GC No competing interests declared, JH consults for Tessera Therapeutics. MF is a co-founder and director of Federation Bio and Viralogic. JW consults for and holds equity in KSQ Therapeutics, Maze Therapeutics, and Tenaya Therapeutics. JSW is a venture partner at 5AM Ventures and a member of the Amgen Scientific Advisory Board. EDAT- 2021/04/28 06:00 MHDA- 2021/10/28 06:00 PMCR- 2021/04/27 CRDT- 2021/04/27 08:39 PHST- 2020/12/16 00:00 [received] PHST- 2021/04/26 00:00 [accepted] PHST- 2021/04/28 06:00 [pubmed] PHST- 2021/10/28 06:00 [medline] PHST- 2021/04/27 08:39 [entrez] PHST- 2021/04/27 00:00 [pmc-release] AID - 65856 [pii] AID - 10.7554/eLife.65856 [doi] PST - epublish SO - Elife. 2021 Apr 27;10:e65856. doi: 10.7554/eLife.65856.