High-throughput tagging of endogenous loci for rapid characterization of protein function

Abstract

To facilitate the interrogation of proteins at scale, we have developed High-throughput Insertion of Tags Across the Genome (HITAG). HITAG enables users to produce libraries of cells, each with a different protein of interest C-terminally tagged, to rapidly characterize protein function. To demonstrate the utility of HITAG, we fused mCherry to a set of 167 stress granule-associated proteins and characterized the factors which drive proteins to strongly accumulate within stress granules.