Efficient opto- and chemogenetic control in a single molecule driven by FRET-modified bioluminescence

Andreas Björefeldt; Jeremy Murphy; Emmanuel L Crespo; Gerard G Lambert; Mansi Prakash; Ebenezer C Ikefuama; Nina Friedman; Tariq M Brown; Diane Lipscombe; Christopher I Moore; Ute Hochgeschwender; Nathan C Shaner

doi:10.1117/1.NPh.11.2.021005

Efficient opto- and chemogenetic control in a single molecule driven by FRET-modified bioluminescence

Neurophotonics. 2024 Apr;11(2):021005. doi: 10.1117/1.NPh.11.2.021005. Epub 2024 Mar 6.

Authors

Andreas Björefeldt^{1

2

3}, Jeremy Murphy², Emmanuel L Crespo^{1

4}, Gerard G Lambert⁵, Mansi Prakash¹, Ebenezer C Ikefuama^{1

6}, Nina Friedman², Tariq M Brown¹, Diane Lipscombe⁷, Christopher I Moore⁷, Ute Hochgeschwender^{1

4

6}, Nathan C Shaner⁵

Affiliations

¹ Central Michigan University, College of Medicine, Mount Pleasant, Michigan, United States.
² Brown University, Department of Neuroscience, Providence, Rhode Island, United States.
³ University of Gothenburg, Institute of Neuroscience and Physiology, Department of Physiology, Gothenburg, Sweden.
⁴ Central Michigan University, Biochemistry, Cell, and Molecular Biology Graduate Program, Mount Pleasant, Michigan, United States.
⁵ University of California, San Diego, School of Medicine, Department of Neurosciences, La Jolla, California, United States.
⁶ Central Michigan University, Neuroscience Graduate Program, Mount Pleasant, Michigan, United States.
⁷ Brown University, Carney Institute for Brain Science, Providence, Rhode Island, United States.

Abstract

Significance: Bioluminescent optogenetics (BL-OG) offers a unique and powerful approach to manipulate neural activity both opto- and chemogenetically using a single actuator molecule (a LuMinOpsin, LMO).

Aim: To further enhance the utility of BL-OG by improving the efficacy of chemogenetic (bioluminescence-driven) LMO activation.

Approach: We developed novel luciferases optimized for Förster resonance energy transfer when fused to the fluorescent protein mNeonGreen, generating bright bioluminescent (BL) emitters spectrally tuned to Volvox Channelrhodopsin 1 (VChR1).

Results: A new LMO generated from this approach (LMO7) showed significantly stronger BL-driven opsin activation compared to previous and other new variants. We extensively benchmarked LMO7 against LMO3 (current standard) and found significantly stronger neuronal activity modulation ex vivo and in vivo, and efficient modulation of behavior.

Conclusions: We report a robust new option for achieving multiple modes of control in a single actuator and a promising engineering strategy for continued improvement of BL-OG.

Keywords: Bioluminescent optogenetics; bioluminescence; chemogenetics; luminopsin; optogenetics.

Abstract

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