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Expanding the Optogenetics Toolkit by Topological Inversion of Rhodopsins.

Cell | 2018

Targeted manipulation of activity in specific populations of neurons is important for investigating the neural circuit basis of behavior. Optogenetic approaches using light-sensitive microbial rhodopsins have permitted manipulations to reach a level of temporal precision that is enabling functional circuit dissection. As demand for more precise perturbations to serve specific experimental goals increases, a palette of opsins with diverse selectivity, kinetics, and spectral properties will be needed. Here, we introduce a novel approach of "topological engineering"-inversion of opsins in the plasma membrane-and demonstrate that it can produce variants with unique functional properties of interest for circuit neuroscience. In one striking example, inversion of a Channelrhodopsin variant converted it from a potent activator into a fast-acting inhibitor that operates as a cation pump. Our findings argue that membrane topology provides a useful orthogonal dimension of protein engineering that immediately permits as much as a doubling of the available toolkit.

Pubmed ID: 30343901 RIS Download

Associated grants

  • Agency: Howard Hughes Medical Institute, United States

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MATLAB (software resource)

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STOCK Gad2tm2(cre)Zjh/J (organism)

RRID:IMSR_JAX:010802

Mus musculus with name STOCK Gad2tm2(cre)Zjh/J from IMSR.

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