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1 Janelia Publications

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    Dudman LabKarpova LabTervo Lab
    10/16/18 | Expanding the optogenetics toolkit by topological inversion of rhodopsins.
    Brown J, Behnam R, Coddington L, Tervo DG, Martin K, Proskurin M, Kuleshova E, Park J, Phillips J, Bergs AC, Gottschalk A, Dudman JT, Karpova AY
    Cell. 2018 Oct 16;175(4):1131-40. doi: 10.1016/j.cell.2018.09.026

    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.

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