Abstract
Fluorescent proteins (FPs) displaying distinct spectra have shed their light on a wide range of biological functions. Moreover, sophisticated biosensors engineered to contain single or multiple FPs, including Förster resonance energy transfer (FRET)-based biosensors, spatiotemporally reveal the molecular mechanisms underlying a variety of pathophysiological processes. However, their usefulness for applied life sciences has yet to be fully explored. Recently, our research group has begun to expand the potential of FPs from basic biological research to the clinic. Here, we describe a method to evaluate the responsiveness of leukemia cells from patients to tyrosine kinase inhibitors using a biosensor based on FP technology and the principle of FRET. Upon phosphorylation of the tyrosine residue of the biosensor, binding of the SH2 domain to phosphotyrosine induces conformational change of the biosensor and brings the donor and acceptor FPs into close proximity. Therefore, kinase activity and response to kinase inhibitors can be monitored by an increase and a decrease in FRET efficiency, respectively. As in basic research, this biosensor resolves hitherto arduous tasks and may provide innovative technological advances in clinical laboratory examinations. State-of-the-art detection devices that enable such innovation are also introduced.
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Acknowledgments
This work was supported in part by Grants-in-Aid from the Japan Society for the Promotion of Science (JSPS; #23659196 and #25670184) and the Adaptable and Seamless Technology Transfer Program through Target-driven R&D from Japan Science and Technology Agency (JST, AS2321227F), as well as grants from the Yasuda Medical Foundation, the Japan Leukemia Research Fund, and the Kato Memorial Bioscience Foundation.
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Fujioka, M., Asano, Y., Nakada, S., Ohba, Y. (2017). SH2 Domain-Based FRET Biosensor for Measuring BCR-ABL Activity in Living CML Cells. In: Machida, K., Liu, B. (eds) SH2 Domains. Methods in Molecular Biology, vol 1555. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6762-9_30
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DOI: https://doi.org/10.1007/978-1-4939-6762-9_30
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Publisher Name: Humana Press, New York, NY
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