Pooled screening for biochemically-defined phenotypes using CRISPuRe-Seq

Phenotypic genetic screening is a powerful approach for biological discovery. CRISPR technologies have enabled routine genome-scale pooled perturbations in cultured cells, creating an opportunity to develop screening methods capable of accessing molecularly meaningful phenotypes. Here we present a generalized strategy, named CRISPuRe-seq, that enables pooled screening of a broad range of protein-based attributes. This approach utilizes the purification of an RNA-barcoded protein-of-interest to identify causative genetic perturbations. As a proof of concept, we screened for modifiers of Interferon alpha signaling and were able to identify all of the primary components of the type 1 Interferon JAK/STAT signaling pathway.