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bioRxiv · 10.1101/2024.01.16.575827

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

Nemcko, F.·Himmelsbach, M.·Loubiere, V.·Yelagandula, R.·Pagani, M.·Brennecke, J.·Elling, U.·Stark, A.·Ameres, S. L.
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Abstract

Determining protein function in a systematic manner 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. Using mouse embryonic stem cells, we showcase ORFtags utility through screens for transcriptional activators, repressors and post-transcriptional regulators. Each screen finds known and novel regulators, including long ORFs not accessible to other methods, revealing that Zfp574 is a highly selective transcriptional activator and that oncogenic fusions frequently function as transactivators.

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Nemcko, F., Himmelsbach, M., Loubiere, V., Yelagandula, R., Pagani, M., Brennecke, J., Elling, U., Stark, A., Ameres, S. L.. 2024-01-17. Proteome-scale tagging and functional screening in mammalian cells by ORFtag. https://doi.org/10.1101/2024.01.16.575827

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