Color-tuned Channelrhodopsins for Multiwavelength Optogenetics

Prigge, Matthias; Schneider, Franziska; Tsunoda, Satoshi P.; Shilyansky, Carrie; Wietek, Jonas; Deisseroth, Karl; Hegemann, Peter

doi:10.1074/jbc.m112.391185

Cited by 156 publications

(159 citation statements)

References 26 publications

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“…In combination with a light-driven bacterial chloride pump (halorhodopsin, NpHR), multiple-color optical activation and silencing of neural circuitry were achieved on the millisecond timescale in living cells and even in freely moving animals (Han and Boyden 2007;Zhang et al 2007). Such light-induced manipulations have been further developed by generating new channelrhodopsin variants (Gunaydin et al 2010;Mattis et al 2012;Prigge et al 2012;Zhang et al 2011), by identification of red-shifted channels in other algae (Govorunova et al 2011;Kianianmomeni et al 2009;Zhang et al 2008) as well as through the improvement of molecular techniques, which allow functional expression in various living animals such as worms, fruit flies, mice and primates (Bi et al 2006;Han et al 2009;Honjo et al 2012;Petreanu et al 2007;Schroll et al 2006;Zhang et al 2007). Through these achievements, optogenetics technology has become widely used in neuroscience as a novel, revolutionary tool for fast neuronal control.…”

Section: Application Of Light-sensitive Modules In Synthetic Biology mentioning

confidence: 99%

Algal photoreceptors: in vivo functions and potential applications

Kianianmomeni

Hallmann

2013

Planta

101

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Section: Application Of Light-sensitive Modules In Synthetic Biology mentioning

confidence: 99%

Algal photoreceptors: in vivo functions and potential applications

Kianianmomeni

Hallmann

2013

Planta

101

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“…Hence, we carefully analyzed the closing kinetics of all three ChRs. PsChR1 and GtACR1 exhibited biphasic off-kinetics comprised of a fast and a slow component like most cation conducting ChRs (15)(16)(17), whereas the decay of GtACR2 was mono-exponential (Fig. 4A).…”

Section: Resultsmentioning

confidence: 99%

Identification of a Natural Green Light Absorbing Chloride Conducting Channelrhodopsin from Proteomonas sulcata

Wietek

Broser

Krause

et al. 2016

Journal of Biological Chemistry

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Chloride conducting channelrhodopsins (ChloCs) are new members of the optogenetic toolbox that enable neuronal inhibition in target cells. Originally, ChloCs have been engineered from cation conducting channelrhodopsins (ChRs), and later identified in a cryptophyte alga genome. We noticed that the sequence of a previously described Proteomonas sulcata ChR (PsChR1) was highly homologous to the naturally occurring and previously reported ChloCs GtACR1/2, but was not recognized as an anion conducting channel. Based on electrophysiological measurements obtained under various ionic conditions, we concluded that the PsChR1 photocurrent at physiological conditions is strongly inward rectifying and predominantly carried by chloride. The maximum activation was noted at excitation with light of 540 nm. An initial spectroscopic characterization of purified protein revealed that the photocycle and the transport mechanism of PsChR1 differ significantly from cation conducting ChRs. Hence, we concluded that PsChR1 is an anion conducting ChR, now renamed PsACR1, with a red-shifted absorption suited for multicolor optogenetic experiments in combination with blue light absorbing cation conducting ChRs.

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“…Through molecular engineering, these tools have been optimized to allow for faster alteration of channels [114,115], response to different wavelengths [116][117][118][119][120], more robust gene expression [114,117,121,122], and channels with stepwise kinetics [117,123,124]. Having tools activated with different wavelengths makes it possible to manipulate the same neuron bidirectionally with both excitatory and inhibitory channels.…”

Section: Optogeneticsmentioning

confidence: 99%

Selective Manipulation of Neural Circuits

Park

Carmel

2016

Neurotherapeutics

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Unraveling the complex network of neural circuits that form the nervous system demands tools that can manipulate specific circuits. The recent evolution of genetic tools to target neural circuits allows an unprecedented precision in elucidating their function. Here we describe two general approaches for achieving circuit specificity. The first uses the genetic identity of a cell, such as a transcription factor unique to a circuit, to drive expression of a molecule that can manipulate cell function. The second uses the spatial connectivity of a circuit to achieve specificity: one genetic element is introduced at the origin of a circuit and the other at its termination. When the two genetic elements combine within a neuron, they can alter its function. These two general approaches can be combined to allow manipulation of neurons with a specific genetic identity by introducing a regulatory gene into the origin or termination of the circuit. We consider the advantages and disadvantages of both these general approaches with regard to specificity and efficacy of the manipulations. We also review the genetic techniques that allow gain-and loss-of-function within specific neural circuits. These approaches introduce light-sensitive channels (optogenetic) or drug sensitive channels (chemogenetic) into neurons that form specific circuits. We compare these tools with others developed for circuit-specific manipulation and describe the advantages of each. Finally, we discuss how these tools might be applied for identification of the neural circuits that mediate behavior and for repair of neural connections.

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Color-tuned Channelrhodopsins for Multiwavelength Optogenetics

Cited by 156 publications

References 26 publications

Algal photoreceptors: in vivo functions and potential applications

Algal photoreceptors: in vivo functions and potential applications

Identification of a Natural Green Light Absorbing Chloride Conducting Channelrhodopsin from Proteomonas sulcata

Selective Manipulation of Neural Circuits

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