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

Michelle S Frei; Samantha A Sanchez; 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:10.1101/2024.02.10.579766

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

bioRxiv [Preprint]. 2024 Feb 11:2024.02.10.579766. doi: 10.1101/2024.02.10.579766.

Authors

Michelle S Frei¹, Samantha A Sanchez^{1

2}, Longwei Liu^{2

3

4}, Falk Schneider^{5

6}, Zichen Wang^{1

7}, Hiroyuki Hakozaki^{1

7}, Yajuan Li², Anne C Lyons^{1

2}, Theresa V Rohm⁸, Jerrold M Olefsky⁸, Lingyan Shi², Johannes Schöneberg^{1

7}, Scott E Fraser^{4

5

9}, Sohum Mehta¹, Yingxiao Wang^{2

3

4}, Jin Zhang^{1

2

7}

Affiliations

¹ Department of Pharmacology, University of California, San Diego, La Jolla, CA, USA.
² Shu Chien-Gene Lay Department of Bioengineering, University of California, San Diego, La Jolla, CA, USA.
³ Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA, USA.
⁴ Alfred E. Mann Department of Biomedical Engineering, University of Southern California, Los Angeles, CA, USA.
⁵ Translational Imaging Center, University of Southern California, Los Angeles, CA, USA.
⁶ Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California, Los Angeles, CA, USA.
⁷ Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.
⁸ Division of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, La Jolla, CA, USA.
⁹ Department of Biological Sciences, Division of Molecular and Computational Biology, University of Southern California, Los Angeles, CA, USA.

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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Preprint

Abstract

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