CRISPR-based functional genomics in human dendritic cells

Jost, M.; Jacobson, A. N.; Hussmann, J. A.; Cirolia, G.; Fischbach, M.; Weissman, J. S.

doi:10.1101/2020.12.22.423985

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bioRxiv · 10.1101/2020.12.22.423985

CRISPR-based functional genomics in human dendritic cells

Jost, M.·Jacobson, A. N.·Hussmann, J. A.·Cirolia, G.·Fischbach, M.·Weissman, J. S.

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Abstract

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.

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Jost, M., Jacobson, A. N., Hussmann, J. A., Cirolia, G., Fischbach, M., Weissman, J. S.. 2020-12-22. CRISPR-based functional genomics in human dendritic cells. https://doi.org/10.1101/2020.12.22.423985

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