title: A genetically targetable near-infrared photosensitizer.
abstract: Upon illumination, photosensitizer molecules produce reactive oxygen species that can be used for functional manipulation of living cells, including protein inactivation, targeted-damage introduction and cellular ablation. Photosensitizers used to date have been either exogenous, resulting in delivery and removal challenges, or genetically encoded proteins that form or bind a native photosensitizing molecule, resulting in a constitutively active photosensitizer inside the cell. We describe a genetically encoded fluorogen-activating protein (FAP) that binds a heavy atom-substituted fluorogenic dye, forming an 'on-demand' activated photosensitizer that produces singlet oxygen and fluorescence when activated with near-infrared light. This targeted and activated photosensitizer (TAPs) approach enables protein inactivation, targeted cell killing and rapid targeted lineage ablation in living larval and adult zebrafish. The near-infrared excitation and emission of this FAP-TAPs provides a new spectral range for photosensitizer proteins that could be useful for imaging, manipulation and cellular ablation deep within living organisms.
authors.affiliation: Department of Chemistry, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.
authors.forename: Jianjun
authors.initials: J
authors.lastname: He
authors.affiliation: Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Developmental Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Missinato
authors.affiliation: Department of Developmental Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Ezenwa
authors.initials: E
authors.lastname: Onuoha
authors.affiliation: Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.
authors.forename: Lydia A
authors.initials: LA
authors.lastname: Perkins
authors.affiliation: Center for Biologic Imaging, Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Simon C
authors.initials: SC
authors.lastname: Watkins
authors.affiliation: Center for Biologic Imaging, Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Claudette M
authors.initials: CM
authors.lastname: St Croix
authors.affiliation: Department of Developmental Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Tsang
authors.affiliation: Department of Chemistry, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.|Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.|Molecular Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.
authors.forename: Marcel P
authors.identifier: http://orcid.org/0000-0002-7370-4848
authors.initials: MP
authors.lastname: Bruchez
mesh_terms: D017209:Apoptosis / Q000502:physiology / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D057809:HEK293 Cells; D006801:Humans; D007259:Infrared Rays; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000737:chemistry* / Q000528:radiation effects*; D011829:Radiation Dosage; D011994:Recombinant Proteins / Q000235:genetics* / Q000627:therapeutic use
chemical_list: D017319:Photosensitizing Agents; D011994:Recombinant Proteins