Human T cells are critical effectors of the immune response as well as autoimmune disease and are attractive tools for cancer immunotherapy. The lab has developed CRISPR-based methods for functional genomic screens identifying genes and networks crucial to the development, fate, and function of human T cells, as well as extending these efforts to other immune cell types.
We developed a method, sgRNA lentiviral infection with Cas9 protein electroporation (SLICE), for genome-wide CRISPR screens in human T cells. We have applied SLICE to discover targets that tune T cell responses to stimulation and immunosuppression and coupled SLICE with single-cell RNA-seq to reveal altered gene expression programs in targeted cells. We recently published a complementary strategy to barcode and track targeted integrations of large DNA templates, enabling pooled knock-in screens in primary human T cells, revealing natural and synthetic sequences that reprogram T cell functions. These technologies allow us to thoroughly investigate the genetic programs that control human immune cell identity and function, adding to our understanding of disease genetics and nominating candidates for therapeutic development.