CRISPR-based perturbation methods for functional analysis of human monocyte-derived dendritic cells

Abstract

Dendritic cells are professional antigen-presenting cells that initiate the adaptive immune response. Their ability to induce effector T cell responses positions them as attractive immunotherapeutic targets. The genes that regulate dendritic cell maturation, migration, and direction of T cell responses are not fully understood. In this thesis, I establish CRISPR-based methods for targeted gene perturbation in human monocyte-derived dendritic cells. I performed a pooled CRISPR knockout screen which identified candidate transcriptional regulators of monocyte-derived dendritic cell maturation in vitro. These methods provide a basis for execution of larger-scale pooled screens and for CRISPR interference-based screens, such as Perturb-seq, in human dendritic cells.