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Optogenetics for light control of biological systems

Abstract

Optogenetic techniques have been developed to allow control over the activity of selected cells within a highly heterogeneous tissue, using a combination of genetic engineering and light. Optogenetics employs natural and engineered photoreceptors, mostly of microbial origin, to be genetically introduced into the cells of interest. As a result, cells that are naturally light-insensitive can be made photosensitive and addressable by illumination and precisely controllable in time and space. The selectivity of expression and subcellular targeting in the host is enabled by applying control elements such as promoters, enhancers and specific targeting sequences to the employed photoreceptor-encoding DNA. This powerful approach allows precise characterization and manipulation of cellular functions and has motivated the development of advanced optical methods for patterned photostimulation. Optogenetics has revolutionized neuroscience during the past 15 years and is primed to have a similar impact in other fields, including cardiology, cell biology and plant sciences. In this Primer, we describe the principles of optogenetics, review the most commonly used optogenetic tools, illumination approaches and scientific applications and discuss the possibilities and limitations associated with optogenetic manipulations across a wide variety of optical techniques, cells, circuits and organisms.

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Fig. 1: Principles of optogenetics.
Fig. 2: The optogenetic actuator toolbox.
Fig. 3: Cell type-specific targeting of optogenetic tools.
Fig. 4: Optical approaches for optogenetic stimulation.
Fig. 5: Expected results in optogenetic experiments.
Fig. 6: Establishing links of causality with optogenetics.
Fig. 7: Optogenetic application for vision restoration, cardiac research and plant modification.

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Acknowledgements

This work was supported by the following funding sources: V.E. is supported by the IHU FOReSIGHT grant (Grant P-ALLOP3-IHU-000), the Axa Research Fund, the European Research Council (ERC) (ERC-2019-AdG; no. 885090) and the Agence National pour la Recherche (ANR-17-CE16-0021). E.E. is supported, in part, by the grants National Institutes of Health (NIH) R01 HL144157, NIH R21EB026152, National Science Foundation (NSF) 1705645 and NSF 1830941. J.V. is supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy (EXC-2049 – 390688087). R.H. is supported by the German Research Foundation (DFG) (Koselleck award HE1640/42-1). P.H. is supported by the DFG (SFB1078, SPP1926, UniSysCat and Neurocure) and the ERC (Stardust 767092). P.H. is Hertie Professor for Neuroscience and supported by the Hertie Foundation. C.L. is supported by the ERC (ERC-2020-AdG, F-addict) and the Swiss National Science Foundations (no. 310030_189188 and CRSII5_186266). M.M. has received funding from the European Union’s Horizon 2020 research and innovation programme under Marie Skłodowska-Curie grant agreement no. 844492. Z.-H.P. is supported by the Ligon Research Center of Vision at Kresge Eye Institute, Dryer Foundation, Herrick Foundation and Research to Prevent Blindness to Department of Ophthalmology, Visual and Anatomical Sciences at Wayne State University School of Medicine. O.Y. is supported by the Joseph and Wolf Lebovic Charitable Foundation Chair for Research in Neuroscience, the ERC (grant no. 819496), the EU Horizon 2020 programme (H2020-ICT-2018-20 DEEPER 101016787) and the Israel Science Foundation (grant no. 3131/20).

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Contributions

Introduction (C.L., E.E., J.V., M.M., O.Y., P.H., R.H., V.E. and Z.-H.P.); Experimentation (J.V., M.M., O.Y., P.H. and V.E.); Results (C.L., E.E., J.V., K.R.K., M.M., O.Y., P.H., R.H., R.R.S., V.E. and Z.-H.P.); Applications (C.L., E.E., K.R.K., O.Y., P.H., R.H., R.R.S., V.E. and Z.-H.P.); Reproducibility and data deposition (C.L., E.E., J.V., K.R.K., M.M., O.Y., P.H., R.H., R.R.S., V.E. and Z.-H.P.); Limitations and optimizations (C.L., E.E., J.V., M.M., O.Y., P.H., R.H., V.E. and Z.-H.P.); Outlook (C.L., E.E., O.Y., P.H., R.H., V.E. and Z.-H.P.); Overview of the Primer (O.Y. and P.H.). Authors are listed in alphabetical order.

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Correspondence to Peter Hegemann.

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Competing interests

Z.-H.P. is a co-inventor on patents related to optogenetic vision restoration and is also a co-founder and scientific advisor of Ray Therapeutics. The other authors declare no competing interests.

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Nature Reviews Methods Primers thanks John Flannery and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Glossary

Microelectrode

An electrode with a micrometre-sized tip used to record single-neuron activity.

Optrode

An electrode coupled to an optical fibre used to record and manipulate neural activity in cells expressing an optogenetic actuator.

Optogenetic actuators

Light-sensitive proteins that transiently modify cellular properties during illumination.

Bidirectional voltage modulation

Changing the voltage in the depolarizing (excitatory) or hyperpolarizing (inhibitory) direction.

Immediate early genes

Genes that are rapidly induced by elevated neural activity such as Fos.

Optical clamp

A technique using light and real-time feedback to keep membrane electrical parameters, such as voltage or action potential shape, at a set desired value.

Antidromic activation

Retrograde propagation of an action potential from the axon to the neuronal soma.

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Emiliani, V., Entcheva, E., Hedrich, R. et al. Optogenetics for light control of biological systems. Nat Rev Methods Primers 2, 55 (2022). https://doi.org/10.1038/s43586-022-00136-4

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