Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins
Open accessAbstract
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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Conventionally used AAV backbones with a 1.3 kb CaMKIIα promoter, WPRE, and human growth hormone polyadenylation signal have a capacity of 2.4 kb.
“Conventionally used AAV backbones which include a commonly used neuronal expression cassette composed of 1.3 kb CaMKIIα promoter (which shows comparable expression efficiency with 0.4 kb CaMKIIα promoter [10] used in CWB), WPRE, and human growth hormone polyadenylation signal [23-25] has a capacity of 2.4 kb.”
From Optimization of AAV expression cassettes to improve packaging capacity and transgene expression in neurons (2014) · cites this paper as [25]
The Results section (‘Inhibiting spikes in pyramidal cells in acute slice’; page not provided) describes an AAV construct controlled by a CaMKIIα promoter. Online Methods (‘Molecular cloning’; page not provided) mentions WPRE, but for lentiviral constructs. Our copy does not give a 1.3 kb promoter length, identify a human growth hormone polyadenylation signal in that AAV backbone, or state a 2.4 kb capacity. (LLM, conf=0.84)
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- Authors
- Mattis, J., Tye, K. M., Ferenczi, E. A., Ramakrishnan, C., O'Shea, D. J., Prakash, R., Gunaydin, L. A., Hyun, M., Fenno, L. E., Gradinaru, V., Yizhar, O., & Deisseroth, K.
- Affiliations
- Stanford University (US)
- Weizmann Institute of Science (IL)
- Howard Hughes Medical Institute (US)
- Corresponding author
- Deisseroth, Karl
- ORCID
- Venue
- Nature Methods
- Published
- 2011-12-18
- Volume
- 9
- Issue
- 2
- Pages
- 159-172
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC4165888
- OpenAlex
- W1969650326
- Cited by
- 780
- References
- 51
- Citations by year
- 2026: 21 · 2025: 41 · 2024: 33 · 2023: 42 · 2022: 61 · 2021: 52 · 2020: 45 · 2019: 55 · 2018: 58 · 2017: 72 · 2016: 66 · 2015: 65 · 2014: 71 · 2013: 75 · 2012: 23
- FWCI
- 22.40
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- CaltechAUTHORS (California Institute of Technology) (submittedVersion)
- Funders
- Howard Hughes Medical Institute
- W. M. Keck Foundation
- McKnight Foundation
- Advanced Research Projects Agency
- National Institute of Mental Health (F32MH088010)
- National Institute of Diabetes and Digestive and Kidney Diseases (DP2DK102256)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Photoreceptor and optogenetics research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neural Engineering
- Keywords
- optogenetics, microbial opsins, figure of merit, light-gated ion channels
- MeSH terms
- Action Potentials, Animals, Kinetics, Light, Pyramidal Cells, Opsins
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