title: Regulation of neural gene transcription by optogenetic inhibition of the RE1-silencing transcription factor.
abstract: Optogenetics provides new ways to activate gene transcription; however, no attempts have been made as yet to modulate mammalian transcription factors. We report the light-mediated regulation of the repressor element 1 (RE1)-silencing transcription factor (REST), a master regulator of neural genes. To tune REST activity, we selected two protein domains that impair REST-DNA binding or recruitment of the cofactor mSin3a. Computational modeling guided the fusion of the inhibitory domains to the light-sensitive Avena sativa light-oxygen-voltage-sensing (LOV) 2-phototrophin 1 (AsLOV2). By expressing AsLOV2 chimeras in Neuro2a cells, we achieved light-dependent modulation of REST target genes that was associated with an improved neural differentiation. In primary neurons, light-mediated REST inhibition increased Na(+)-channel 1.2 and brain-derived neurotrophic factor transcription and boosted Na(+) currents and neuronal firing. This optogenetic approach allows the coordinated expression of a cluster of genes impinging on neuronal activity, providing a tool for studying neuronal physiology and correcting gene expression changes taking place in brain diseases.
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy; Department of Experimental Medicine, University of Genova, 16132 Genova, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Paonessa
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy; Department of Experimental Medicine, University of Genova, 16132 Genova, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Criscuolo
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy;
authors.forename: Silvio
authors.initials: S
authors.lastname: Sacchetti
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy;
authors.forename: Davide
authors.initials: D
authors.lastname: Amoroso
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy;
authors.forename: Helena
authors.initials: H
authors.lastname: Scarongella
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy; Department of Experimental Medicine, University of Genova, 16132 Genova, Italy.
authors.forename: Federico
authors.initials: F
authors.lastname: Pecoraro Bisogni
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy;
authors.forename: Emanuele
authors.initials: E
authors.lastname: Carminati
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy;
authors.forename: Giacomo
authors.initials: G
authors.lastname: Pruzzo
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy;
authors.forename: Luca
authors.initials: L
authors.lastname: Maragliano
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy; fabrizia.cesca@iit.it fabio.benfenati@iit.it.
authors.forename: Fabrizia
authors.initials: F
authors.lastname: Cesca
authors.affiliation: Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16132 Genova, Italy; Department of Experimental Medicine, University of Genova, 16132 Genova, Italy fabrizia.cesca@iit.it fabio.benfenati@iit.it.
authors.forename: Fabio
authors.initials: F
authors.lastname: Benfenati
mesh_terms: D000818:Animals; D018554:Avena / Q000235:genetics; D045744:Cell Line, Tumor; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D051379:Mice; D009474:Neurons / Q000378:metabolism*; D062308:Optogenetics / Q000379:methods*; D010940:Plant Proteins / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic; D014161:Transduction, Genetic; D000099229:RE1-Silencing Transcription Factor
chemical_list: D002843:Chromatin; D004247:DNA; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D000099229:RE1-Silencing Transcription Factor
keywords: AsLOV2; REST/NRSF; gene transcription; molecular dynamics; optogenetics