Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States; Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States; California Institute for Quantitative Biosciences, University of California, San Francisco, San Francisco, United States; Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, United States
Amy N Jacobson
Department of Bioengineering, Stanford University, Stanford, United States; ChEM-H, Stanford University, Stanford, United States
Jeffrey A Hussmann
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States; Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States; California Institute for Quantitative Biosciences, University of California, San Francisco, San Francisco, United States; Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, United States; Whitehead Institute for Biomedical Research, Cambridge, United States
Giana Cirolia
Chan Zuckerberg Biohub, San Francisco, United States
Michael A Fischbach
Department of Bioengineering, Stanford University, Stanford, United States; ChEM-H, Stanford University, Stanford, United States; Chan Zuckerberg Biohub, San Francisco, United States
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States; Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, United States; California Institute for Quantitative Biosciences, University of California, San Francisco, San Francisco, United States; Whitehead Institute for Biomedical Research, Cambridge, United States; Department of Biology, Massachusetts Institute of Technology, Cambridge, United States
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.
