Details

Autor(en) / Beteiligte

• Kleinlogel, Sonja
• Terpitz, Ulrich
• Legrum, Barbara
• Gökbuget, Deniz
• Boyden, Edward S
• Bamann, Christian
• Wood, Phillip G
• Bamberg, Ernst

Titel

A gene-fusion strategy for stoichiometric and co-localized expression of light-gated membrane proteins

Ist Teil von

• Nature methods, 2011-12, Vol.8 (12), p.1083-1088

Ort / Verlag

United States

Erscheinungsjahr

2011

Quelle

MEDLINE

Beschreibungen/Notizen

• The precise co-localization and stoichiometric expression of two different light-gated membrane proteins can vastly improve the physiological usefulness of optogenetics for the modulation of cell excitability with light. Here we present a gene-fusion strategy for the stable 1:1 expression of any two microbial rhodopsins in a single polypeptide chain. By joining the excitatory channelrhodopsin-2 with the inhibitory ion pumps halorhodopsin or bacteriorhodopsin, we demonstrate light-regulated quantitative bi-directional control of the membrane potential in HEK293 cells and neurons in vitro. We also present synergistic rhodopsin combinations of channelrhodopsin-2 with Volvox carteri channelrhodopsin-1 or slow channelrhodopsin-2 mutants, to achieve enhanced spectral or kinetic properties, respectively. Finally, we demonstrate the utility of our fusion strategy to determine ion-turnovers of as yet uncharacterized rhodopsins, exemplified for archaerhodopsin and CatCh, or to correct pump cycles, exemplified for halorhodopsin.