TY  - JOUR
AU  - Spreen, Anika
AU  - Sadanandan, Nidish Ponath
AU  - Schneider, Martin Winfried
AU  - Kuehn, Enrico
AU  - Leemisa, Andries Napo
AU  - De Zio, Roberta
AU  - Meyer, Niklas
AU  - Busse, Wayne
AU  - Erlinghagen, Bela
AU  - Adenauer, Lea
AU  - Opitz, Thoralf
AU  - Bönigk, Wolfgang
AU  - Schiewer, Enrico
AU  - Heer, Jonas
AU  - Schmitz, Dietmar
AU  - Vierock, Johannes
AU  - Beck, Heinz
AU  - Hegemann, Peter
AU  - Baier, Herwig
AU  - Schneider-Warme, Franziska
AU  - Sierra, Yinth Andrea Bernal
AU  - Seifert, Reinhard
TI  - Optogenetic silencing by combining a rhodopsin cyclase with an engineered cGMP-gated potassium channel.
JO  - Science advances
VL  - 11
IS  - 48
SN  - 2375-2548
CY  - Washington, DC [u.a.]
PB  - Assoc.
M1  - DZNE-2025-01325
SP  - eadx1195
PY  - 2025
AB  - 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.
KW  - Optogenetics: methods
KW  - Animals
KW  - Cyclic GMP: metabolism
KW  - Mice
KW  - Rhodopsin: metabolism
KW  - Rhodopsin: genetics
KW  - Humans
KW  - Hippocampus: metabolism
KW  - Hippocampus: cytology
KW  - Guanylate Cyclase: metabolism
KW  - Guanylate Cyclase: genetics
KW  - Zebrafish
KW  - Potassium Channels: metabolism
KW  - Potassium Channels: genetics
KW  - Protein Engineering
KW  - Myocytes, Cardiac: metabolism
KW  - Neurons: metabolism
KW  - HEK293 Cells
KW  - Cyclic GMP (NLM Chemicals)
KW  - Rhodopsin (NLM Chemicals)
KW  - Guanylate Cyclase (NLM Chemicals)
KW  - Potassium Channels (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:41313760
C2  - pmc:PMC12662207
DO  - DOI:10.1126/sciadv.adx1195
UR  - https://pub.dzne.de/record/282562
ER  -