Design of growth-coupled measurements of transcription factor function

This article presents a proposal for measuring the function of transcription factors (TFs) using a growth-coupled assay integrated with a gene circuit. This innovative approach links TF activity to bacterial cell growth, allowing for quantitative assessments of TF function across various families, starting with RamR, Lacl, and TetR. The methodology involves using plasmids designed for specific TFs, employing a range of calibration variants to span their activity dynamic range. The project aims to generate comprehensive datasets that will enhance understanding of TF repression mechanisms and the impact of mutations on their function, ultimately facilitating advancements in synthetic biology and therapeutic applications. The research is supported by a collaborative team from institutions such as NIST and Harvard Medical School, and it outlines a structured plan for both pilot and large-scale data collection efforts to expand the characterization of TFs across different families.