Details

Autor(en) / Beteiligte

• Mukund, Adi X.
• Tycko, Josh
• Allen, Sage J.
• Robinson, Stephanie A.
• Andrews, Cecelia
• Sinha, Joydeb
• Ludwig, Connor H.
• Spees, Kaitlyn
• Bassik, Michael C.
• Bintu, Lacramioara

Titel

High-throughput functional characterization of combinations of transcriptional activators and repressors

Ist Teil von

• Cell systems, 2023-09, Vol.14 (9), p.746-763.e5

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2023

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• 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 recruitment to target genes by using a small-molecule drug. Altogether, we outline the basic principles of how effector domains combine to regulate gene expression and demonstrate their value in building precise and flexible synthetic biology tools. A record of this paper’s transparent peer review process is included in the supplemental information. [Display omitted] •We screen 8,400 effector domain pairs for transcriptional activation and repression•We find synergy between weak activators and antagonism between strong activators•We find that repressors tend to dominate activators when co-recruited•We build a TF that changes from activator to repressor upon adding a small molecule Mukund et al. measure transcriptional activation and repression for thousands of transcriptional effector domain pairs using a high-throughput reporter assay in human cells. This study helps outline the basic principles of how effector domains can be combined to build novel synthetic biology tools for precise and flexible gene regulation.