Engineering aspects of bioluminescence resonance energy transfer systems

Abstract

Development of optical probes for sensing biological functions in vivo is in high demand in modern biology. With several inherent advantages, the bioluminescence resonance energy transfer (BRET)-based sensors are rapidly expanding and showing great utilities in the study of protein-protein interactions (PPIs), protein dimerization, signal transduction, etc. Since its inception in the late nineties, BRET-related research has gained significant momentum in terms of adding versatility to the assay format and wider applicability where it has been suitably used. Beyond the scope of quantitative measurement of PPIs and protein dimerization, molecular imaging applications based on BRET assays have broadened its scope for screening pharmacologically important compounds by in vivo imaging and high-throughput screening (HTS) methods. Taking examples from literatures, this chapter will contribute to an in-depth knowledge on engineering requirements of BRET components such as donor, acceptor, substrate chemistry, and instrumentations. BRET applications having significant contributions toward making it an attractive single-format assay are also discussed.