CRISPR-based functional genomics in human dendritic cells

• Main Authors: Marco Jost, Amy N Jacobson, Jeffrey A Hussmann, Giana Cirolia, Michael A Fischbach, Jonathan S Weissman
• Format: Article
• Language: English
• Published: eLife Sciences Publications Ltd 2021-04-01
• Series: eLife
• Subjects: CRISPR; dendritic cells; bacteroides thetaiotaomicron; functional genomics; inter-individual variation
• Online Access: https://elifesciences.org/articles/65856

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.