The present palette of opsin-based optogenetic tools lacks a light-gated potassium (K<sup>+</sup>) channel desirable for silencing of excitable cells. Here, we describe the construction of a blue-light-induced K<sup>+</sup> channel 1 (BLINK1) engineered by fusing the plant LOV2-Jα photosensory module to the small viral K<sup>+</sup> channel Kcv. BLINK1 exhibits biophysical features of Kcv, including K<sup>+</sup> selectivity and high single-channel conductance, but reversibly photoactivates in blue light. Opening of BLINK1 channels hyperpolarizes the cell to the K<sup>+</sup> equilibrium potential. Ectopic expression of BLINK1 reversibly inhibits the escape response in light-exposed zebrafish larvae. BLINK1 therefore provides a single-component optogenetic tool that can establish prolonged, physiological hyperpolarization of cells at low light intensities.
Engineering of a light-gated potassium channel / C. Cosentino, L. Alberio, S. Gazzarrini, M. Aquila, E. Romano, S. Cermenati, P. Zuccolini, J. Petersen, M. Beltrame, J.L. Van Etten, J.M. Christie, G. Thiel, A. Moroni. - In: SCIENCE. - ISSN 0036-8075. - 348:6235(2015 May 08), pp. 707-710. [10.1126/science.aaa2787]
Engineering of a light-gated potassium channel
C. CosentinoPrimo
;L. AlberioSecondo
;S. Gazzarrini;M. Aquila;S. Cermenati;M. Beltrame;A. MoroniUltimo
2015
Abstract
The present palette of opsin-based optogenetic tools lacks a light-gated potassium (K+) channel desirable for silencing of excitable cells. Here, we describe the construction of a blue-light-induced K+ channel 1 (BLINK1) engineered by fusing the plant LOV2-Jα photosensory module to the small viral K+ channel Kcv. BLINK1 exhibits biophysical features of Kcv, including K+ selectivity and high single-channel conductance, but reversibly photoactivates in blue light. Opening of BLINK1 channels hyperpolarizes the cell to the K+ equilibrium potential. Ectopic expression of BLINK1 reversibly inhibits the escape response in light-exposed zebrafish larvae. BLINK1 therefore provides a single-component optogenetic tool that can establish prolonged, physiological hyperpolarization of cells at low light intensities.File | Dimensione | Formato | |
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