Genetically Encoded Whole Cell Biosensor for Drug Discovery of HIF-1 Interaction Inhibitors
Artikel i vetenskaplig tidskrift, 2022

The heterodimeric transcription factor, hypoxia inducible factor-1 (HIF-1), is an important anticancer target as it supports the adaptation and response of tumors to hypoxia. Here, we optimized the repressed transactivator yeast two-hybrid system to further develop it as part of a versatile yeast-based drug discovery platform and validated it using HIF-1. We demonstrate both fluorescence-based and auxotrophy-based selections that could detect HIF-1α/HIF-1β dimerization inhibition. The engineered genetic selection is tunable and able to differentiate between strong and weak interactions, shows a large dynamic range, and is stable over different growth phases. Furthermore, we engineered mechanisms to control for cellular activity and off-target drug effects. We thoroughly characterized all parts of the biosensor system and argue this tool will be generally applicable to a wide array of protein-protein interaction targets. We anticipate this biosensor will be useful as part of a drug discovery platform, particularly when screening DNA-encoded new modality drugs.

biosensor

high-throughput screen

genetic circuit

new modality

drug discovery

directed evolution

Författare

Louis Scott

AstraZeneca AB

Chalmers, Biologi och bioteknik, Systembiologi

Mark J. Wigglesworth

AstraZeneca AB

Verena Siewers

Chalmers, Biologi och bioteknik, Systembiologi

Andy M. Davis

AstraZeneca AB

Florian David

Chalmers, Biologi och bioteknik, Systembiologi

ACS Synthetic Biology

2161-5063 (eISSN)

Vol. 11 10 3182-3189

Ämneskategorier

Farmaceutisk vetenskap

Biokemi och molekylärbiologi

Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

DOI

10.1021/acssynbio.2c00274

PubMed

36223492

Mer information

Senast uppdaterat

2024-03-07