Genetically targeted fluorogenic macromolecules for subcellular imaging and cellular perturbation

• Published in: Biomaterials Vol. 66; pp. 1 - 8
• Main Authors: Magenau, Andrew J.D, Saurabh, Saumya, Andreko, Susan K, Telmer, Cheryl A, Schmidt, Brigitte F, Waggoner, Alan S, Bruchez, Marcel P
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.10.2015
• Subjects: Advanced Basic Science; Atom transfer radical polymerization (ATRP); Binding; Biomaterial structure-property relationships; Cellular; Cellular structure; Dentistry; Fluorescent Dyes - chemistry; Fluorescent Dyes - pharmacokinetics; Fluorogen-activating proteins (FAPs); Gene Targeting - methods; Genetically targetable macromolecules; HeLa Cells; Humans; Macromolecules; Medical services; Microscopy, Fluorescence - methods; Molecular Imaging - methods; Polymer modified cells; Polymerization; Polymers; Protein polymer hybrids; Proteins; Proteins - chemistry; Proteins - genetics; Proteins - pharmacokinetics; Subcellular Fractions - metabolism; Subcellular Fractions - ultrastructure; Subcellular imaging; Targeting polymers to subcellular organelles; Protein polymer hybrids; Targeting polymers to subcellular organelles; Polymer modified cells; Subcellular imaging; Atom transfer radical polymerization (ATRP); Biomaterial structure-property relationships; Genetically targetable macromolecules; Fluorogen-activating proteins (FAPs)
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2015.07.002

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.