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The RAS family of proteins is amongst the most highly mutated in human cancers and has so far eluded drug therapy. Currently, much effort is being made to discover mutant RAS inhibitors and in vitro screening for RAS-binding drugs must be followed by cell-based assays. Here, we have developed a robust set of bioluminescence resonance energy transfer (BRET)-based RAS biosensors that enable monitoring of RAS-effector interaction inhibition in living cells. These include KRAS, HRAS and NRAS and a variety of different mutations that mirror those found in human cancers with the major RAS effectors such as CRAF, PI3K and RALGDS. We highlighted the utility of these RAS biosensors by showing a RAS-binding compound is a potent pan-RAS-effector interactions inhibitor in cells. The RAS biosensors represent a useful tool to investigate and characterize the potency of anti-RAS inhibitors in cells and more generally any RAS protein-protein interaction (PPI) in cells.

Original publication

DOI

10.7554/eLife.37122

Type

Journal article

Journal

Elife

Publication Date

10/07/2018

Volume

7

Keywords

BRET, RAS, biosensors, cancer, cancer biology, cell-based assays, drug development, human, intracellular antibody, mouse, protein-protein interaction inhibition, small molecule inhibitors, Bioluminescence Resonance Energy Transfer Techniques, Biosensing Techniques, Energy Transfer, HEK293 Cells, Humans, Mutation, Protein Binding, Protein Interaction Domains and Motifs, Proto-Oncogene Proteins p21(ras), Signal Transduction, Small Molecule Libraries