title: Optogenetics. 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.
authors.affiliation: Department of Biosciences, University of Milano, Italy.
authors.forename: Cristian
authors.initials: C
authors.lastname: Cosentino
authors.affiliation: Department of Biosciences, University of Milano, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Alberio
authors.affiliation: Department of Biosciences, University of Milano, Italy.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Gazzarrini
authors.affiliation: Department of Biosciences, University of Milano, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Aquila
authors.affiliation: Department of Biosciences, University of Milano, Italy.
authors.forename: Edoardo
authors.initials: E
authors.lastname: Romano
authors.affiliation: Department of Biosciences, University of Milano, Italy.
authors.forename: Solei
authors.initials: S
authors.lastname: Cermenati
authors.affiliation: Department of Biosciences, University of Milano, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Zuccolini
authors.affiliation: Institute of Molecular, Cell and Systems Biology, University of Glasgow, UK.
authors.forename: Jan
authors.initials: J
authors.lastname: Petersen
authors.affiliation: Department of Biosciences, University of Milano, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Beltrame
authors.affiliation: Department of Plant Pathology and Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, NE 68583-0900, USA.
authors.forename: James L
authors.initials: JL
authors.lastname: Van Etten
authors.affiliation: Institute of Molecular, Cell and Systems Biology, University of Glasgow, UK.
authors.forename: John M
authors.initials: JM
authors.lastname: Christie
authors.affiliation: Membrane Biophysics, Technical University of Darmstadt, Darmstadt, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Thiel
authors.affiliation: Department of Biosciences, University of Milano, Italy. anna.moroni@unimi.it.
authors.forename: Anna
authors.initials: A
authors.lastname: Moroni
mesh_terms: D000818:Animals; D018554:Avena / Q000378:metabolism; D055592:Biophysical Phenomena; D057809:HEK293 Cells; D006801:Humans; D007814:Larva; D008027:Light; D062308:Optogenetics; D056949:Phototropins / Q000737:chemistry / Q000235:genetics; D015221:Potassium Channels / Q000737:chemistry / Q000235:genetics; D011487:Protein Conformation / Q000528:radiation effects; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000528:radiation effects*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics; D015027:Zebrafish
chemical_list: C405463:Kcv potassium channel, Chlorella virus; D056949:Phototropins; D015221:Potassium Channels; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C000598723:blue-light-induced K channel 1