RT Journal Article
SR Electronic
T1 CRISPR-based functional genomics in human dendritic cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.12.22.423985
DO 10.1101/2020.12.22.423985
A1 Jost, Marco
A1 Jacobson, Amy N.
A1 Hussmann, Jeffrey A.
A1 Cirolia, Giana
A1 Fischbach, Michael A.
A1 Weissman, Jonathan S.
YR 2020
UL http://biorxiv.org/content/early/2020/12/22/2020.12.22.423985.abstract
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.