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Molecular Optimization of Rhodopsin-Based Tools for Neuroscience Applications

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Rhodopsin

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2501))

Abstract

There is no question that genetically encoded tools have revolutionized neuroscience. These include optically modulated tools for writing-in (optogenetics) and reading-out (calcium, voltage, and neurotransmitter indicators) neural activity as well as precision expression of these reagents using virally mediated delivery. With few exceptions, these powerful approaches are derived from naturally occurring molecules that are sourced from diverse organisms that span all kingdoms of life. Successful expression of genetic tools in standard neuroscience model organisms requires optimizing gene structure, taking into account differences in both protein translation and trafficking. Myriad approaches have resolved these two challenges, resulting in order-of-magnitude increases in functional expression. In this chapter, we focus on synthesizing prior experience in successfully enabling the transition of genes across kingdoms with a goal of facilitating the production of the next generation of molecular tools for neuroscience. We then provide a detailed protocol that allows expression and testing of novel genetically encoded tools in mammalian cell lines and primary cultured neurons.

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Author information

Authors and Affiliations

  1. Departments of Psychiatry and Neuroscience, University of Texas Austin Dell Medical School, Austin, TX, USA

    Lief E. Fenno

  2. Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel

    Rivka Levy & Ofer Yizhar

Authors
  1. Lief E. Fenno
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  2. Rivka Levy
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  3. Ofer Yizhar
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Corresponding author

Correspondence to Ofer Yizhar .

Editor information

Editors and Affiliations

  1. Institut de Biologie Structurale (IBS), Université Grenoble Alpes, CEA, CNRS, Grenoble, France

    Valentin Gordeliy

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Fenno, L.E., Levy, R., Yizhar, O. (2022). Molecular Optimization of Rhodopsin-Based Tools for Neuroscience Applications. In: Gordeliy, V. (eds) Rhodopsin. Methods in Molecular Biology, vol 2501. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2329-9_14

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  • DOI: https://doi.org/10.1007/978-1-0716-2329-9_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2328-2

  • Online ISBN: 978-1-0716-2329-9

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