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Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins

Mattis, Joanna and Tye, Kay M. and Ferenczi, Emily A. and Ramakrishnan, Charu and O’Shea, Daniel J. and Prakash, Rohit and Gunaydin, Lisa A. and Hyun, Minsuk and Fenno, Lief E. and Gradinaru, Viviana and Yizhar, Ofer and Deisseroth, Karl (2012) Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins. Nature Methods, 9 (2). pp. 159-172. ISSN 1548-7091. PMCID PMC4165888. http://resolver.caltech.edu/CaltechAUTHORS:20121206-140040487

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Abstract

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.


Item Type:Article
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URLURL TypeDescription
http://dx.doi.org/10.1038/NMETH.1808 DOIArticle
http://www.nature.com/nmeth/journal/v9/n2/full/nmeth.1808.htmlPublisherArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4165888/PubMed CentralArticle
http://rdcu.be/mSyHPublisherFree ReadCube access
ORCID:
AuthorORCID
Gradinaru, Viviana0000-0001-5868-348X
Additional Information:© 2012 Nature Publishing Group. Received 09 May 2011; Accepted 10 November 2011; Published online 18 December 2011; Corrected online 10 January 2012. 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. Author Contributions: J.M., K.M.T., E.A.F., C.R., R.P., O.Y. and K.D. contributed to study design and data interpretation. J.M. coordinated all experiments and data analysis. J.M., K.M.T., E.A.F., D.J.O., R.P. and L.E.F. contributed to acquisition of electrophysiological data. C.R. cloned all constructs, cultured primary neurons, performed transfections and managed viral packaging processes. D.J.O. wrote custom analysis scripts and analyzed all electrophysiological data. M.H. contributed to data analysis. J.M., K.M.T., C.R., L.A.G. and V.G. contributed to the histological processing and fluorescence imaging. K.D. supervised all aspects of the work. J.M., K.M.T., E.A.F. and K.D. wrote the paper.
Funders:
Funding AgencyGrant Number
BioX undergraduate summer research programUNSPECIFIED
Stanford Medical Scientist Training ProgramUNSPECIFIED
NIH1F32MH088010-01
International Fulbright Science and Technology AwardUNSPECIFIED
Stanford Graduate FellowshipUNSPECIFIED
National Institute on Drug AbuseUNSPECIFIED
National Institute of Neurological Disorders and Stroke (NINDS)UNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Defense Advanced Research Projects Agency (DARPA)UNSPECIFIED
W. M. Keck FoundationUNSPECIFIED
McKnight Foundation UNSPECIFIED
Gatsby Charitable FoundationUNSPECIFIED
National Institute of Mental Health (NIMH)UNSPECIFIED
Subject Keywords:Neuroscience
PubMed Central ID:PMC4165888
Record Number:CaltechAUTHORS:20121206-140040487
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20121206-140040487
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:35861
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:07 Dec 2012 21:44
Last Modified:22 Nov 2016 22:59

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