PT  - JOURNAL ARTICLE
AU  - Jost, Marco
AU  - Jacobson, Amy N.
AU  - Hussmann, Jeffrey A.
AU  - Cirolia, Giana
AU  - Fischbach, Michael A.
AU  - Weissman, Jonathan S.
TI  - CRISPR-based functional genomics in human dendritic cells
AID  - 10.1101/2020.12.22.423985
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.12.22.423985
4099  - http://biorxiv.org/content/early/2020/12/22/2020.12.22.423985.short
4100  - http://biorxiv.org/content/early/2020/12/22/2020.12.22.423985.full
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 >93% 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 genes that control this specificity, highlighting the potential of our method to pinpoint determinants of inter-individual variation in immune responses. 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.