Enhancing the Efficacy, Utility, and Throughput of the Transcription Block Survival Peptide Library Screening Platform.

ABSTRACT: Genetically encoded peptide library screening is a powerful strategy for discovering inhibitors of protein-protein and protein-DNA interactions. The Transcription Block Survival (TBS) assay enables the in vivo selection of peptides that antagonize transcription factor (TF) function by linking the inhibition of DNA binding to E. coli survival. However, previous TBS implementations required laborious re-engineering of the mDHFR coding region for each new target, limiting utility. Here, we present an enhanced and streamlined TBS platform that increases throughput, simplifies target switching, and improves selection stringency. By relocating TF DNA-binding sites from within the mDHFR coding sequence into the mDHFR 5'-promoter/untranslated region, we preserve mDHFR folding and fun

SUBMITTER: Brennan A 

PROVIDER: S-EPMC12648313 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature