Optogenetic silencing by combining a rhodopsin cyclase with an engineered cGMP-gated potassium channel
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<sup>+</sup> channel), which pairs the rhodopsin-guanylyl cyclase CaRhGC with customized SthK K<sup>+</sup> 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<sup>+</sup>-based cell silencing.
Authors
- Jonas Heer (ORCID: https://orcid.org/0009-0003-1844-5331)
- Martin Schneider (ORCID: https://orcid.org/0000-0001-9760-6292)
- Enrico Schiewer (ORCID: https://orcid.org/0000-0001-7913-5597)
- Anika Spreen (ORCID: https://orcid.org/0009-0002-5649-4364)
- W. F. Busse (ORCID: https://orcid.org/0009-0006-4364-7914)
- Franziska Schneider‐Warme (ORCID: https://orcid.org/0000-0002-5873-9436)
- Peter Hegemann (ORCID: https://orcid.org/0000-0003-3589-6452)
- Johannes Vierock (ORCID: https://orcid.org/0000-0001-7368-5539)
- Nils Meyer (ORCID: https://orcid.org/0009-0004-6794-1267)
- Herwig Baier (ORCID: https://orcid.org/0000-0002-7268-0469)
- Roberta De Zio (ORCID: https://orcid.org/0000-0002-1777-2088)
- Heinz Beck (ORCID: https://orcid.org/0000-0002-8961-998X)
- Nidish Ponath Sadanandan (ORCID: https://orcid.org/0000-0003-1534-831X)
- Yinth Andrea Bernal Sierra (ORCID: https://orcid.org/0000-0003-3067-7496)
- Enrico Kuehn (ORCID: https://orcid.org/0000-0003-3428-5860)
- Wolfgang Bönigk (ORCID: https://orcid.org/0000-0003-0217-9086)
- Dietmar Schmitz (ORCID: https://orcid.org/0000-0003-2741-5241)
- Thoralf Opitz (ORCID: https://orcid.org/0009-0001-2419-5004)
- Andries Napo Leemisa (ORCID: https://orcid.org/0009-0009-8156-2155)
- Reinhard Seifert (ORCID: https://orcid.org/0009-0003-5872-4845)
- Bela Erlinghagen (ORCID: https://orcid.org/0009-0002-3198-3137)
- Lea Adenauer
Institutions
- University of Bonn (DE)
- University of Freiburg (DE)
- Center of Advanced European Studies and Research (DE)
- University Medical Center Freiburg (DE)
- German Center for Neurodegenerative Diseases (DE)
- Humboldt-Universität zu Berlin (DE)
- Max Planck Institute of Neurobiology (DE)
- Max Planck Institute of Biochemistry (DE)
- Bernstein Center for Computational Neuroscience Berlin (DE)
- University of Bari Aldo Moro (IT)
- Charité - Universitätsmedizin Berlin (DE)
Publication Details
- Journal
- Science Advances
- Published
- 2025-11-28
- DOI
- https://doi.org/10.1126/sciadv.adx1195
- Primary Topic
- Photoreceptor and optogenetics research
- Type
- article
- Field-Weighted Citation Impact
- 0.00