Optogenetic silencing by combining a rhodopsin cyclase with an engineered cGMP-gated potassium channel
Year: 2025
Type: article
Abstract: Since the advent of optogenetics, great progress has been made in developing tools to modulate and detect cellular activity using light. We present a two-component optogenetic silencing tool, RoCK (rhodopsin cyclase/K + channel), which pairs the rhodopsin-guanylyl cyclase CaRhGC with customized SthK K + channels that are engineered to open selectively upon guanosine 3′,5′-monophosphate (cGMP) binding. By enhancing the cGMP sensitivity and open probability of SthK mutants, we obtained four channel variants suited for different levels of cGMP concentration. CaRhGC’s membrane-bound nature enables localized cGMP production, and the lack of dark activity reduces the risk for off-target effects. Optimized RoCK effectively modulated cellular activity in mouse hippocampal neurons, in acute hippocampal slices, and in rabbit cardiomyocytes. In zebrafish, RoCK silenced motor neurons in vivo, suppressing the characteristic coiling behavior of embryos, thus highlighting its potential for behavioral studies. In summary, RoCK expands our optogenetic toolkit threefold for fast cGMP production, fast cGMP sensing, and K + -based cell silencing.
Source: Science Advances
Authors: Anika Spreen, Nidish Ponath Sadanandan, Martin Winfried Schneider, Enrico Kuehn, Andries Napo Leemisa, Roberta De Zio, Niklas Meyer, Wayne Busse, Bela Erlinghagen, Lea Adenauer, Thoralf Opitz, Wolfgang Bönigk, Enrico Schiewer, Jonas Heer, Dietmar Schmitz, Johannes Vierock, Heinz Beck, Peter Hegemann, Herwig Baier, Franziska Schneider-Warme, Yinth Andrea Bernal Sierra, Reinhard Seifert
Institutions: Humboldt-Universität zu Berlin, Max Planck Institute for Neurobiology of Behavior – caesar, Max Planck Institute for Biological Intelligence, University of Freiburg, University Medical Center Freiburg, Universitäts-Herzzentrum Freiburg-Bad Krozingen, University of Bari Aldo Moro, Charité - Universitätsmedizin Berlin, University of Bonn, German Center for Neurodegenerative Diseases, Bernstein Center for Computational Neuroscience Berlin, NeuroCure Cluster of Excellence, Freie Universität Berlin
Language: English
Subfield: Cellular and Molecular Neuroscience
Field: Neuroscience
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $4,500
Awards: G1717570230, G2707994935, Optische Steuerung von zellbiophysikalischen Prozessen in nativem Herzgewebe: Beleuchtung der spezifischen Einflüsse von interstitiellen und myeloiden Zellen auf die kardiale Elektrophysiologie, Entwicklung und Anwendung von RoCK, einem neuen Rhodopsinzyklase/K+ Kanalbasierten optogenetischen Werkzeug zur Inhibierung erregbarer Zellen, G6662086760, SFB 1425: Die heterozelluläre Natur kardialer Läsionen: Identitäten, Interaktionen, Implikationen
OpenAlex ID: W4416801234