A genome-wide CRISPR Screen identifies targets that drive Tolerogenic Dendritic Cells

However, the key molecular pathways driving their differentiation and tolerogenic phenotype remain elusive. Here we investigated the molecular mechanisms underlying the functional status of TolDCs in response to different tolerizing agents and identified CD86 as a consistent and robust marker downregulated in tolerogenic state.

CD86 is functionally required for TolDC-mediated suppression of T cell proliferation and cytokine secretion, supporting its use as a surrogate marker of tolerogenic potential. Leveraging CD86 expression as a readout, we performed a genome-wide CRISPR-Cas9 knockout screen and identified regulators of TolDC-associated phenotypes. This approach uncovered UBE2L6 as a novel regulator of CD86 expression through modulation of an ISG15-USP18-associated pathway, implicating its involvement in tolerogenic functions.