title: Genetically targeted fluorogenic macromolecules for subcellular imaging and cellular perturbation.
abstract: The alteration of cellular functions by anchoring macromolecules to specified organelles may reveal a new area of therapeutic potential and clinical treatment. In this work, a unique phenotype was evoked by influencing cellular behavior through the modification of subcellular structures with genetically targetable macromolecules. These fluorogen-functionalized polymers, prepared via controlled radical polymerization, were capable of exclusively decorating actin, cytoplasmic, or nuclear compartments of living cells expressing localized fluorgen-activating proteins. The macromolecular fluorogens were optimized by establishing critical polymer architecture-biophysical property relationships which impacted binding rates, binding affinities, and the level of internalization. Specific labeling of subcellular structures was realized at nanomolar concentrations of polymer, in the absence of membrane permeabilization or transduction domains, and fluorogen-modified polymers were found to bind to protein intact after delivery to the cytosol. Cellular motility was found to be dependent on binding of macromolecular fluorogens to actin structures causing rapid cellular ruffling without migration.
authors.affiliation: Molecular Biosensors and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA; Department of Chemistry, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA. Electronic address: amagenau@coe.drexel.edu.
authors.forename: Andrew J D
authors.initials: AJ
authors.lastname: Magenau
authors.affiliation: Department of Chemistry, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA.
authors.forename: Saumya
authors.initials: S
authors.lastname: Saurabh
authors.affiliation: Molecular Biosensors and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA.
authors.forename: Susan K
authors.initials: SK
authors.lastname: Andreko
authors.affiliation: Molecular Biosensors and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA; Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA.
authors.forename: Cheryl A
authors.initials: CA
authors.lastname: Telmer
authors.affiliation: Molecular Biosensors and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA.
authors.forename: Brigitte F
authors.initials: BF
authors.lastname: Schmidt
authors.affiliation: Molecular Biosensors and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA; Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA.
authors.forename: Alan S
authors.initials: AS
authors.lastname: Waggoner
authors.affiliation: Molecular Biosensors and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA; Department of Chemistry, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA; Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA. Electronic address: bruchez@cmu.edu.
authors.forename: Marcel P
authors.initials: MP
authors.lastname: Bruchez
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry / Q000493:pharmacokinetics*; D018390:Gene Targeting / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000493:pharmacokinetics; D013347:Subcellular Fractions / Q000378:metabolism* / Q000648:ultrastructure*
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins
keywords: Atom transfer radical polymerization (ATRP); Biomaterial structure-property relationships; Fluorogen-activating proteins (FAPs); Genetically targetable macromolecules; Polymer modified cells; Protein polymer hybrids; Subcellular imaging; Targeting polymers to subcellular organelles