Potassium channel-based optogenetic silencing

• Published in: Nature communications Vol. 9; no. 1; pp. 4611 - 13
• Main Authors: Bernal Sierra, Yinth Andrea, Rost, Benjamin R., Pofahl, Martin, Fernandes, António Miguel, Kopton, Ramona A., Moser, Sylvain, Holtkamp, Dominik, Masala, Nicola, Beed, Prateep, Tukker, John J., Oldani, Silvia, Bönigk, Wolfgang, Kohl, Peter, Baier, Herwig, Schneider-Warme, Franziska, Hegemann, Peter, Beck, Heinz, Seifert, Reinhard, Schmitz, Dietmar
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 05.11.2018; Nature Publishing Group; Nature Portfolio
• Subjects: 14/69; 42; 631/1647/2253; 631/378; 64/116; 64/60; 692/4019; 9/74; 96/95; Adenylyl Cyclases - genetics; Adenylyl Cyclases - metabolism; Adenylyl Cyclases - radiation effects; Animals; Animals, Genetically Modified; Biological activity; Cardiomyocytes; Channelrhodopsins - radiation effects; Gene Expression - genetics; Gene Expression - radiation effects; Genetics; HEK293 Cells; Humanities and Social Sciences; Humans; Hyperpolarization; Information processing; Light; Luminous intensity; Mice; Models, Animal; multidisciplinary; Myocytes, Cardiac - metabolism; Neurons - metabolism; Neurons - radiation effects; Optics; Optogenetics - methods; Potassium; Potassium Channels - genetics; Potassium Channels - metabolism; Potassium Channels - radiation effects; Regulatory sequences; Rhodopsin - pharmacology; Science; Science (multidisciplinary); Silencer Elements, Transcriptional; Temporal resolution; Zebrafish
• DOI: 10.1038/s41467-018-07038-8
• ISSN: 2041-1723

Optogenetics enables manipulation of biological processes with light at high spatio-temporal resolution to control the behavior of cells, networks, or even whole animals. In contrast to the performance of excitatory rhodopsins, the effectiveness of inhibitory optogenetic tools is still insufficient. Here we report a two-component optical silencer system comprising photoactivated adenylyl cyclases (PACs) and the small cyclic nucleotide-gated potassium channel SthK. Activation of this ‘PAC-K’ silencer by brief pulses of low-intensity blue light causes robust and reversible silencing of cardiomyocyte excitation and neuronal firing. In vivo expression of PAC-K in mouse and zebrafish neurons is well tolerated, where blue light inhibits neuronal activity and blocks motor responses. In combination with red-light absorbing channelrhodopsins, the distinct action spectra of PACs allow independent bimodal control of neuronal activity. PAC-K represents a reliable optogenetic silencer with intrinsic amplification for sustained potassium-mediated hyperpolarization, conferring high operational light sensitivity to the cells of interest. Optogenetic tools enable precise experimental control of the behaviour of cells. Here, the authors introduce a genetically-encoded two-protein system that enables silencing of excitable cells such as neurons and cardiomyocytes using blue light, and demonstrate its utility both in vitro and In vivo.