In vivo expression of a light-activatable potassium channel using unnatural amino acids.
Author: Kang JY, Kawaguchi D, Coin I, Xiang Z, O'Leary DD, Slesinger PA, Wang L.
Affiliation:
The Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Conference/Journal: Neuron.
Date published: 2013 Oct 16
Other:
Volume ID: 80 , Issue ID: 2 , Pages: 358-70 , Special Notes: doi: 10.1016/j.neuron.2013.08.016 , Word Count: 167
Optical control of protein function provides excellent spatial-temporal resolution for studying proteins in situ. Although light-sensitive exogenous proteins and ligands have been used to manipulate neuronal activity, a method for optical control of neuronal proteins using unnatural amino acids (Uaa) in vivo is lacking. Here, we describe the genetic incorporation of a photoreactive Uaa into the pore of an inwardly rectifying potassium channel Kir2.1. The Uaa occluded the pore, rendering the channel nonconducting, and, on brief light illumination, was released to permit outward K(+) current. Expression of this photoinducible inwardly rectifying potassium (PIRK) channel in rat hippocampal neurons created a light-activatable PIRK switch for suppressing neuronal firing. We also expanded the genetic code of mammals to express PIRK channels in embryonic mouse neocortex in vivo and demonstrated a light-activated PIRK current in cortical neurons. These principles could be generally expanded to other proteins expressed in the brain to enable optical regulation.
Copyright © 2013 Elsevier Inc. All rights reserved.
PMID: 24139041 [PubMed - indexed for MEDLINE] PMCID: PMC3815458 [Available on 2014/10/16]
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