CRISPR-based functional genomics in human dendritic cells

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.

Interrogating immune cells and cancer with CRISPR-Cas9

“…In summary, these methods for CRISPR screening in human T cells present exciting opportunities to understand gene regulation in primary cells. Recent reports of CRISPR editing in primary human B cells and monocyte derived-dendritic cells may further extend the possibilities of human immune cell CRISPR screens [43][44][45]. Furthermore, future screens may increasingly focus on complex immune phenotypes beyond those of proliferation or stimulation; high-dimensional readouts will be particularly useful in studying complex cell states (Box 2).…”