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@ARTICLE{Spreen:282562,
author = {Spreen, Anika and Sadanandan, Nidish Ponath and Schneider,
Martin Winfried and Kuehn, Enrico and Leemisa, Andries Napo
and De Zio, Roberta and Meyer, Niklas and Busse, Wayne and
Erlinghagen, Bela and Adenauer, Lea and Opitz, Thoralf and
Bönigk, Wolfgang and Schiewer, Enrico and Heer, Jonas and
Schmitz, Dietmar and Vierock, Johannes and Beck, Heinz and
Hegemann, Peter and Baier, Herwig and Schneider-Warme,
Franziska and Sierra, Yinth Andrea Bernal and Seifert,
Reinhard},
title = {{O}ptogenetic silencing by combining a rhodopsin cyclase
with an engineered c{GMP}-gated potassium channel.},
journal = {Science advances},
volume = {11},
number = {48},
issn = {2375-2548},
address = {Washington, DC [u.a.]},
publisher = {Assoc.},
reportid = {DZNE-2025-01325},
pages = {eadx1195},
year = {2025},
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.},
keywords = {Optogenetics: methods / Animals / Cyclic GMP: metabolism /
Mice / Rhodopsin: metabolism / Rhodopsin: genetics / Humans
/ Hippocampus: metabolism / Hippocampus: cytology /
Guanylate Cyclase: metabolism / Guanylate Cyclase: genetics
/ Zebrafish / Potassium Channels: metabolism / Potassium
Channels: genetics / Protein Engineering / Myocytes,
Cardiac: metabolism / Neurons: metabolism / HEK293 Cells /
Cyclic GMP (NLM Chemicals) / Rhodopsin (NLM Chemicals) /
Guanylate Cyclase (NLM Chemicals) / Potassium Channels (NLM
Chemicals)},
cin = {AG Schmitz / Bonn common},
ddc = {500},
cid = {I:(DE-2719)1810004 / I:(DE-2719)6000011},
pnm = {351 - Brain Function (POF4-351) / 899 - ohne Topic
(POF4-899)},
pid = {G:(DE-HGF)POF4-351 / G:(DE-HGF)POF4-899},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:41313760},
pmc = {pmc:PMC12662207},
doi = {10.1126/sciadv.adx1195},
url = {https://pub.dzne.de/record/282562},
}