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Green-Sensitive, Long-Lived, Step-Functional Anion Channelrhodopsin-2 Variant as a High-Potential Neural Silencing Tool.
Green-Sensitive, Long-Lived, Step-Functional Anion Channelrhodopsin-2 Variant as a High-Potential Neural Silencing Tool.
- Source :
-
The journal of physical chemistry letters [J Phys Chem Lett] 2020 Aug 06; Vol. 11 (15), pp. 6214-6218. Date of Electronic Publication: 2020 Jul 22. - Publication Year :
- 2020
-
Abstract
- Anion channelrhodopsin-2 (GtACR2) was identified from the alga Guillardia theta as a light-gated anion channel, providing a powerful neural silencing tool for optogenetics. To expand its molecular properties, we produced here GtACR2 variants by strategic mutations on the four residues around the retinal chromophore (i.e., R129, G152, P204, and C233). After the screening with the Escherichia coli expression system, we estimated spectral sensitivities and the anion channeling function by using the HEK293 expression system. Among the mutants, triple (R129M/G152S/C233A) and quadruple (R129M/G152S/P204T/C233A) mutants showed the significantly red-shifted absorption maxima (λ <subscript>max</subscript> = 498 and 514 nm, respectively) and the long-lived channel-conducting states (the half-life times were 3.4 and 5.4 s, respectively). In addition, both mutants can be activated and inactivated by different wavelengths, representing their step-functional ability. We nicknamed the quadruple mutant "GLaS-ACR2" from its green-sensitive, long-lived, step-functional properties. The unique characteristics of GLaS-ACR2 suggest its high potential as a neural silencing tool.
- Subjects :
- Anions chemistry
Channelrhodopsins genetics
Cryptophyta genetics
Escherichia coli genetics
Gene Expression Regulation
Green Chemistry Technology
HEK293 Cells
Humans
Ion Transport
Mutation
Optogenetics
Photochemical Processes
Channelrhodopsins chemistry
Cryptophyta chemistry
Fluorescent Dyes chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1948-7185
- Volume :
- 11
- Issue :
- 15
- Database :
- MEDLINE
- Journal :
- The journal of physical chemistry letters
- Publication Type :
- Academic Journal
- Accession number :
- 32697087
- Full Text :
- https://doi.org/10.1021/acs.jpclett.0c01406