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

ABSTRACT: The systematic determination of protein function is a key goal of modern biology, but remains challenging with current approaches. Here, we present ORFtag, a versatile, cost-effective and highly efficient method for the massively parallel tagging and functional interrogation of proteins at proteome scale. ORFtag utilizes retroviral vectors bearing a promoter, peptide tag and splice donor to generate fusions between the tag and endogenous ORFs. We demonstrate the utility of ORFtag through functional screens for transcriptional activators, repressors and post-transcriptional regulators in mouse embryonic stem cells. Each screen finds known and novel regulators, including long ORFs inaccessible by other methods, revealing that Zfp574 is a highly selective transcriptional activator and that oncogenic fusions often function as transactivators.

ORGANISM(S): Mus musculus

PROVIDER: GSE267110 | GEO | 2024/05/10

REPOSITORIES: GEO

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