Unmet Need: Biosensors are a widely used tool for tracking spatiotemporal dynamics of molecular activities. However, many biosensors are based on the Förster resonance energy transfer (FRET) between fluorescent proteins (FPs), and a major drawback has been their limited multiplexing capability due to the broad emission spectra of FPs and the availability of spectral space, as well as the lack of standards for calibrating the signal over the long term. Disclosed herein are novel barcoding libraries and biosensor calibration standards that enable multiplexed long-term tracking of biomolecular signals in 2D and 3D cultures.
Value Proposition:
· Novel biosensor barcoding for multiplexed spatiotemporal profiling of biomolecular signals in cells
· FRET-compatible and standardized to enable measurement across image settings
· Broad use applications in research and clinical diagnostics and drug screening
Technology Overview: Researchers at Johns Hopkins University have developed novel FRET-based biosensor barcoding methodology capable of simultaneously tracking large numbers of biomolecular activities in live cells over extended periods of time. Briefly, researchers created a FRET-compatible barcoding library of FPs and calibration standards capable of normalizing signals across various imaging settings. Cells expressing biosensors are separately labeled with barcoded proteins, and imaged for spatiotemporal molecular profiling. Multiplexed biosensor imaging can facilitate comprehensive identification and kinetic characterization of multiple cellular activities. Broadly applicable, utilities include use in cellular assays for drug screening or detecting signaling irregularities for improved patient diagnostics and management.
Stage of Development: Proof of concept validation completed
Publication:
1. Yang JM, Chi WY, Liang J, Takayanagi S, Iglesias P, Huang CH. Deciphering cell signaling networks with massively multiplexed biosensor barcoding. Cell. 2021; 184(25), 6193–6
2. Wang S, Chi WY, Au G, Huang CC, Yang JM, Huang CH. Chi WY, Au G, Liang J, Chen CC, Huang CH, Yang JM. Imaging and analysis for simultaneous tracking of fluorescent biosensors in barcoded cells. STAR Protocols. 2022; 3(3), 101611.
3. Reconstructing signaling networks using biosensor barcoding. Methods Mol Biol. 2024;2800:189–202. 206.e14.
