Diverse optogenetic tools have allowed versatile control over neural activity. Many depolarizing and hyperpolarizing tools have now been developed in multiple laboratories and tested across different preparations, presenting opportunities but also making it difficult to draw direct comparisons. This challenge has been compounded by the dependence of performance on parameters such as vector, promoter, expression time, illumination, cell type and many other variables. As a result, it has become increasingly complicated for end users to select the optimal reagents for their experimental needs. For a rapidly growing field, critical figures of merit should be formalized both to establish a framework for further development and so that end users can readily understand how these standardized parameters translate into performance. Here we systematically compared microbial opsins under matched experimental conditions to extract essential principles and identify key parameters for the conduct, design and interpretation of experiments involving optogenetic techniques.
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We thank A. Andalman, T. Davidson, I. Diester, S. Evans, I. Goshen, D. Mattis, I. Witten, L. Grosenick, S.-Y. Kim and C. Perry for helpful discussions, M. Lin (Stanford University) for ChIEF clones, and all memebers of the Deisseroth laboratory for their support. All viruses were packaged at University of North Carolina Vector Core. Supported by Bio-X and the Stanford Medical Scientist Training Program (J.M.), the US National Institute of Mental Health (1F32MH088010-01, K.M.T.), and the International Fulbright Science and Technology Award and a Stanford Graduate Fellowship (E.A.F.). K.D. is supported by National Institute of Mental Health, National Institute on Drug Abuse, National Institute of Neurological Disorders and Stroke, Howard Hughes Medical Institute, the Defense Advanced Research Projects Agency Reorganization and Plasticity to Accelerate Injury Recovery Program, the Keck Foundation, the McKnight Foundation and the Gatsby Charitable Foundation.
The authors declare no competing financial interests.
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Mattis, J., Tye, K., Ferenczi, E. et al. Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins. Nat Methods 9, 159–172 (2012). https://doi.org/10.1038/nmeth.1808
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