Skretas Lab has been leveraging the fluorescence of Escherichia coli bacteria engineered to express fusions of misfolding-prone (MisP) human proteins with green fluorescent protein (GFP) as a high-throughput screening readout for identifying chemical compounds that rescue disease-associated protein misfolding and aggregation.

In a newly published study in ACS Synthetic Biology, the team defined conditions that optimize the performance of this assay. Using MisP variants of p53, a protein linked to cancer, as a model system, they demonstrated that robust overexpression conditions significantly improve the assay’s dynamic range and sensitivity. These optimized conditions were also effective for other MisPs, such as amyloid-β peptide (Aβ) and Cu/Zn superoxide dismutase variants, which are associated with Alzheimer’s disease and amyotrophic lateral sclerosis (ALS), respectively.

These insights provide a valuable framework for the standardization of MisP-GFP screening assays, supporting their broader application in the discovery of therapeutic agents targeting protein misfolding and aggregation.

Congratulations to the team!

For more information, please read the full article here.