title: Optogenetic Inhibitor of the Transcription Factor CREB.
abstract: Current approaches for optogenetic control of transcription do not mimic the activity of endogenous transcription factors, which act at numerous sites in the genome in a complex interplay with other factors. Optogenetic control of dominant negative versions of endogenous transcription factors provides a mechanism for mimicking the natural regulation of gene expression. Here we describe opto-DN-CREB, a blue-light-controlled inhibitor of the transcription factor CREB created by fusing the dominant negative inhibitor A-CREB to photoactive yellow protein (PYP). A light-driven conformational change in PYP prevents coiled-coil formation between A-CREB and CREB, thereby activating CREB. Optogenetic control of CREB function was characterized in vitro, in HEK293T cells, and in neurons where blue light enabled control of expression of the CREB targets NR4A2 and c-Fos. Dominant negative inhibitors exist for numerous transcription factors; linking these to optogenetic domains offers a general approach for spatiotemporal control of native transcriptional events.
authors.affiliation: Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S 3H6, Canada; Department of Medicinal Chemistry, Faculty of Pharmacy, Assiut University, Assiut 71515, Egypt.
authors.forename: Ahmed M
authors.initials: AM
authors.lastname: Ali
authors.affiliation: Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S 3H6, Canada.
authors.forename: Jakeb M
authors.initials: JM
authors.lastname: Reis
authors.affiliation: Department of Molecular Biosciences, and Center for Computational Biology, University of Kansas, Lawrence, KS 66045, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Xia
authors.affiliation: Program in Neurosciences & Mental Health at The Hospital for Sick Children, and Departments of Psychology and Physiology and the Institute of Medical Science, University of Toronto, Toronto, ON M5G 1X8, Canada.
authors.forename: Asim J
authors.initials: AJ
authors.lastname: Rashid
authors.affiliation: Program in Neurosciences & Mental Health at The Hospital for Sick Children, and Departments of Psychology and Physiology and the Institute of Medical Science, University of Toronto, Toronto, ON M5G 1X8, Canada.
authors.forename: Valentina
authors.initials: V
authors.lastname: Mercaldo
authors.affiliation: Program in Neurosciences & Mental Health at The Hospital for Sick Children, and Departments of Psychology and Physiology and the Institute of Medical Science, University of Toronto, Toronto, ON M5G 1X8, Canada.
authors.forename: Brandon J
authors.initials: BJ
authors.lastname: Walters
authors.affiliation: Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S 3H6, Canada.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Brechun
authors.affiliation: Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S 3H6, Canada.
authors.forename: Vitali
authors.initials: V
authors.lastname: Borisenko
authors.affiliation: Program in Neurosciences & Mental Health at The Hospital for Sick Children, and Departments of Psychology and Physiology and the Institute of Medical Science, University of Toronto, Toronto, ON M5G 1X8, Canada.
authors.forename: Sheena A
authors.initials: SA
authors.lastname: Josselyn
authors.affiliation: Department of Molecular Biosciences, and Center for Computational Biology, University of Kansas, Lawrence, KS 66045, USA.
authors.forename: John
authors.initials: J
authors.lastname: Karanicolas
authors.affiliation: Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S 3H6, Canada. Electronic address: awoolley@chem.utoronto.ca.
authors.forename: G Andrew
authors.initials: GA
authors.lastname: Woolley
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D050882:CREB-Binding Protein / Q000037:antagonists & inhibitors* / Q000378:metabolism; D003373:Coumaric Acids / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D057809:HEK293 Cells; D006801:Humans; D008027:Light; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2 / Q000378:metabolism; D062308:Optogenetics; D020283:Photoreceptors, Microbial / Q000737:chemistry / Q000378:metabolism; D011422:Propionates; D011485:Protein Binding; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D003373:Coumaric Acids; C539882:NR4A2 protein, human; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D020283:Photoreceptors, Microbial; D011422:Propionates; D016760:Proto-Oncogene Proteins c-fos; C050162:photoactive yellow protein, Bacteria; D004247:DNA; D050882:CREB-Binding Protein; C494363:CREBBP protein, human; C495469:p-coumaric acid
keywords: CREB; PYP; dominant negative; endogenous; genetically encoded; optogenetics; photo-control; photoactive yellow protein; photoisomerization; transcription factor