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Science & Technology

Improved Fluorescent Protein for Enhanced Utilization of FRET Biosensors

By 22nd May 2020No Comments

The following study was conducted by Scientists from University of California San Diego, USA; The George Washington University, Washington, USA; University of California San Diego, La Jolla, USA. Study is published in Nature Communications Journal as detailed below.

Nature Communications; Volume 11, Article Number: 1848; (2020)

A Rationally Enhanced Red Fluorescent Protein Expands the Utility of FRET Biosensors                                                                       


Genetically encoded Förster Resonance Energy Transfer (FRET)-based biosensors are powerful tools to illuminate spatiotemporal regulation of cell signaling in living cells, but the utility of the red spectrum for biosensing was limited due to a lack of bright and stable red fluorescent proteins. Here, we rationally improve the photophysical characteristics of the coral-derived fluorescent protein TagRFP-T. We show that a new single-residue mutant, super-TagRFP (stagRFP) has nearly twice the molecular brightness of TagRFP-T and negligible photoactivation. stagRFP facilitates significant improvements on multiple green-red biosensors as a FRET acceptor and is an efficient FRET donor that supports red/far-red FRET biosensing. Capitalizing on the ability of stagRFP to couple with multiple FRET partners, we develop a novel multiplex method to examine the confluence of signaling activities from three kinases simultaneously in single living cells, providing evidence for a role of Src family kinases in regulating growth factor induced Akt and ERK activities.


Nature Communications



Mo, G. C. H., C. Posner, et al. (2020). “A rationally enhanced red fluorescent protein expands the utility of FRET biosensors.” Nature Communications 11(1): 1848.