Genome_wide_screens_to_find_regulators_of_STAT_proteins___

Next-generation sequencing studies have vastly improved over the last decade and have been widely implemented in clinical practice, particularly to identify genetic variations found within a population. However, the vast majority of these genetic variants in human disease genes are interpreted as variants of uncertain significance (VUSs), since their effects on the gene function and disease pathogenesis are unknown. Thus, there is a dire need for scaled functional interpretation of VUSs. Saturation genome editing (SGE) uses CRISPR-Cas9 genome editing to introduce all possible variants into a targeted region of the native genomic locus, through homology-directed repair. Hence, all variants are coupled to a functional readout, where genotype is linked to the phenotype, following DNA sequencing to quantify the functional effect of each variant. In this project, we propose to use SGE to map the functional consequences of all genetic variants in candidate genes involved in cytokine signaling cascades, with variants associated with immune diseases and previously identified to regulate T cell activation. Specifically, we aim to: profile the molecular phenotypes of all gene variants; and explore the effects of the variants on gene function. Altogether, this project will provide a relevant assessment of variants affecting genes linked to a spectrum of immune phenotypes.