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Engineering rhodopsins' activation spectra using a FRET-based approach.

Publication ,  Journal Article
Beck, C; Gong, Y
Published in: Biophysical journal
May 2022

In the past decade, optogenetics has become a nearly ubiquitous tool in neuroscience because it enables researchers to manipulate neural activity with high temporal resolution and genetic specificity. Rational engineering of optogenetic tools has produced channelrhodopsins with a wide range of kinetics and photocurrent magnitude. Genome mining for previously unidentified species of rhodopsin has uncovered optogenetic tools with diverse spectral sensitivities. However, rational engineering of a rhodopsin has thus far been unable to re-engineer spectral sensitivity while preserving full photocurrent. Here, we developed and characterized ChroME-mTFP, a rhodopsin-fluorescent protein fusion that drives photocurrent through Förster resonance energy transfer (FRET). This FRET-opsin mechanism artificially broadened the activation spectrum of the blue-green-light-activated rhodopsin ChroME by approximately 50 nm, driving higher photocurrent at blue-shifted excitation wavelengths without sacrificing kinetics. The excitation spectra's increase at short wavelengths enabled us to optogenetically excite neurons at lower excitation powers with shorter wavelengths of light. Increasing this rhodopsin's sensitivity to shorter, bluer wavelengths pushes it toward dual-channel, crosstalk-free optogenetic stimulation and imaging with green-light-activated sensors. However, this iteration of FRET-opsin suffers from some imaging-light-induced photocurrent crosstalk from green or yellow light due to maintained, low-efficiency excitation at longer wavelengths.

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Published In

Biophysical journal

DOI

EISSN

1542-0086

ISSN

0006-3495

Publication Date

May 2022

Volume

121

Issue

9

Start / End Page

1765 / 1776

Related Subject Headings

  • Rhodopsin
  • Optogenetics
  • Opsins
  • Fluorescence Resonance Energy Transfer
  • Channelrhodopsins
  • Biophysics
  • 51 Physical sciences
  • 34 Chemical sciences
  • 31 Biological sciences
  • 06 Biological Sciences
 

Citation

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Beck, C., & Gong, Y. (2022). Engineering rhodopsins' activation spectra using a FRET-based approach. Biophysical Journal, 121(9), 1765–1776. https://doi.org/10.1016/j.bpj.2022.03.024
Beck, Connor, and Yiyang Gong. “Engineering rhodopsins' activation spectra using a FRET-based approach.Biophysical Journal 121, no. 9 (May 2022): 1765–76. https://doi.org/10.1016/j.bpj.2022.03.024.
Beck C, Gong Y. Engineering rhodopsins' activation spectra using a FRET-based approach. Biophysical journal. 2022 May;121(9):1765–76.
Beck, Connor, and Yiyang Gong. “Engineering rhodopsins' activation spectra using a FRET-based approach.Biophysical Journal, vol. 121, no. 9, May 2022, pp. 1765–76. Epmc, doi:10.1016/j.bpj.2022.03.024.
Beck C, Gong Y. Engineering rhodopsins' activation spectra using a FRET-based approach. Biophysical journal. 2022 May;121(9):1765–1776.
Journal cover image

Published In

Biophysical journal

DOI

EISSN

1542-0086

ISSN

0006-3495

Publication Date

May 2022

Volume

121

Issue

9

Start / End Page

1765 / 1776

Related Subject Headings

  • Rhodopsin
  • Optogenetics
  • Opsins
  • Fluorescence Resonance Energy Transfer
  • Channelrhodopsins
  • Biophysics
  • 51 Physical sciences
  • 34 Chemical sciences
  • 31 Biological sciences
  • 06 Biological Sciences