Engineering of a light-gated potassium channel

Cristian Cosentino; Laura Alberio; Sabrina Gazzarrini; Marco Aquila; Edoardo Romano; Solei Cermenati; Paolo Zuccolini; Jan Petersen; Monica Beltrame; James L. Van Etten; John M. Christie; Gerhard Thiel; Anna Moroni

doi:10.1126/science.aaa2787

Engineering of a light-gated potassium channel

Cristian Cosentino, Laura Alberio, Sabrina Gazzarrini, Marco Aquila, Edoardo Romano, Solei Cermenati, Paolo Zuccolini, Jan Petersen, Monica Beltrame, James L. Van Etten, John M. Christie, Gerhard Thiel, Anna Moroni

Plant Pathology

Research output: Contribution to journal › Article › peer-review

110 Scopus citations

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.

Original language	English (US)
Pages (from-to)	707-710
Number of pages	4
Journal	Science
Volume	348
Issue number	6235
DOIs	https://doi.org/10.1126/science.aaa2787
State	Published - May 8 2015

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title = "Engineering of a light-gated potassium channel",

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.",

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AU - Cosentino, Cristian

AU - Alberio, Laura

AU - Gazzarrini, Sabrina

AU - Aquila, Marco

AU - Romano, Edoardo

AU - Cermenati, Solei

AU - Zuccolini, Paolo

AU - Petersen, Jan

AU - Beltrame, Monica

AU - Van Etten, James L.

AU - Christie, John M.

AU - Thiel, Gerhard

AU - Moroni, Anna

PY - 2015/5/8

Y1 - 2015/5/8

N2 - 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.

AB - 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.

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Engineering of a light-gated potassium channel

Abstract

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