biostudies-literatureMukund AXNIDDK NIH HHSNHGRI NIH HHSNIGMS NIH HHS746-763.e5https://www.ebi.ac.uk/biostudies/studies/S-EPMC10642976biostudies-literatureUnknown14(9)Despite growing knowledge of the functions of individual human transcriptional effector domains, much less is understood about how multiple effector domains within the same protein combine to regulate gene expression. Here, we measure transcriptional activity for 8,400 effector domain combinations by recruiting them to reporter genes in human cells. In our assay, weak and moderate activation domains synergize to drive strong gene expression, whereas combining strong activators often results in weaker activation. In contrast, repressors combine linearly and produce full gene silencing, and repressor domains often overpower activation domains. We use this information to build a synthetic transcription factor whose function can be tuned between repression and activation independent of recruitCell systemsHigh-throughput functional characterization of combinations of transcriptional activators and repressors.PMC10642976T32 GM145402R01 HG011866R35 GM128947T32 GM007365F99 DK126120K00 DK126120T32 GM113854Mukund AXSpees KRobinson SATycko JAllen SJBintu LAndrews CLudwig CHSinha JBassik MCfalseHigh-throughput functional characterization of combinations of transcriptional activators and repressors.Despite growing knowledge of the functions of individual human transcriptional effector domains, much less is understood about how multiple effector domains within the same protein combine to regulate gene expression. Here, we measure transcriptional activity for 8,400 effector domain combinations by recruiting them to reporter genes in human cells. In our assay, weak and moderate activation domains synergize to drive strong gene expression, whereas combining strong activators often results in weaker activation. In contrast, repressors combine linearly and produce full gene silencing, and repressor domains often overpower activation domains. We use this information to build a synthetic transcription factor whose function can be tuned between repression and activation independent of recruit2023-01-01T00:00:00Z2023 Sep2026-06-02T06:32:51.469Z2025-04-06T16:10:59.408ZS-EPMC106429763754303910.1016/j.cels.2023.07.001