Proteome-scale tagging and functional screening in mammalian cells by ORFtag [PRO-Seq]

Determining protein function in a systematic and comprehensive manner is a key goal in modern molecular biology research but is challenging with current approaches. Here, we introduce ORFtag, a versatile method for the massively parallel protein tagging and interrogation of protein function at the proteome scale with minimal effort and expense. We demonstrate the versatility of ORFtag in mouse and human cells with a series of functional screens for transcriptional activators, repressors and post-transcriptional gene regulatory proteins. Overall design: PRO-Seq for Zfp574-depleted (AID) mESC