Far-red chemigenetic biosensors for multi-dimensional and super-resolved kinase activity imaging

Michelle S. Frei; Samantha A. Sanchez; Xinchang He; Longwei Liu; Falk Schneider; Zichen Wang; Hiroyuki Hakozaki; Yajuan Li; Anne C. Lyons; Theresa V. Rohm; Jerrold M. Olefsky; Lingyan Shi; Johannes Schöneberg; Scott E. Fraser; Sohum Mehta; Yingxiao Wang; Jin Zhang

doi:10.1038/s41587-025-02642-8

BioengineeringNature BiotechnologyApr 21, 2025

Peer-reviewed journal article

Far-red chemigenetic biosensors for multi-dimensional and super-resolved kinase activity imaging

Michelle S. Frei iD, Samantha A. Sanchez iD, Xinchang He iD, Longwei Liu iD, Falk Schneider iD, Zichen Wang iD, Hiroyuki Hakozaki iD, Yajuan Li iD, Anne C. Lyons iD, Theresa V. Rohm iD, Jerrold M. Olefsky iD, Lingyan Shi iD, Johannes Schöneberg iD, Scott E. Fraser iD, Sohum Mehta iD, Yingxiao Wang iD, Jin Zhang iD

Abstract

Fluorescent biosensors revolutionized biomedical science by enabling the direct measurement of signaling activities in living cells, yet the current technology is limited in resolution and dimensionality. Here, we introduce highly sensitive chemigenetic kinase activity biosensors that combine the genetically encodable self-labeling protein tag HaloTag7 with bright far-red-emitting synthetic fluorophores. This technology enables five-color biosensor multiplexing, 4D activity imaging, and functional super-resolution imaging via stimulated emission depletion (STED) microscopy.

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