title: Polymer beacons for luminescence and magnetic resonance imaging of DNA delivery.
abstract: The delivery of nucleic acids with polycations offers tremendous potential for developing highly specific treatments for various therapeutic targets. Although materials have been developed and studied for polynucleotide transfer, the biological mechanisms and fate of the synthetic vehicle has remained elusive due to the limitations with current labeling technologies. Here, we have developed polymer beacons that allow the delivery of nucleic acids to be visualized at different biological scales. The polycations have been designed to contain repeated oligoethyleneamines, for binding and compacting nucleic acids into nanoparticles, and lanthanide (Ln) chelates [either luminescent europium (Eu(3+)) or paramagnetic gadolinium (Gd(3+))]. The chelated Lns allow the visualization of the delivery vehicle both on the nm/microm scale via microscopy and on the sub-mm scale via MRI. We demonstrate that these delivery beacons effectively bind and compact plasmid (p)DNA into nanoparticles and protect nucleic acids from nuclease damage. These delivery beacons efficiently deliver pDNA into cultured cells and do not exhibit toxicity. Micrographs of cultured cells exposed to the nanoparticle complexes formed with fluorescein-labeled pDNA and the europium-chelated polymers reveal effective intracellular imaging of the delivery process. MRI of bulk cells exposed to the complexes formulated with pDNA and the gadolinium-chelated structures show bright image contrast, allowing visualization of effective intracellular delivery on the tissue-scale. Because of their versatility, these delivery beacons posses remarkable potential for tracking and understanding nucleic acid transfer in vitro, and have promise as in vivo theranostic agents.
authors.affiliation: Department of Chemistry, Macromolecules and Interfaces Institute, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
authors.forename: Joshua M
authors.initials: JM
authors.lastname: Bryson
authors.forename: Katye M
authors.initials: KM
authors.lastname: Fichter
authors.forename: Wen-Jang
authors.initials: WJ
authors.lastname: Chu
authors.forename: Jing-Huei
authors.initials: JH
authors.lastname: Lee
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Louis A
authors.initials: LA
authors.lastname: Madsen
authors.forename: Patrick M
authors.initials: PM
authors.lastname: McLendon
authors.forename: Theresa M
authors.initials: TM
authors.lastname: Reineke
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000235:genetics; D005063:Europium / Q000737:chemistry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005682:Gadolinium / Q000737:chemistry; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D007089:Image Enhancement / Q000379:methods; D028581:Lanthanoid Series Elements / Q000737:chemistry; D049449:Luminescence; D008279:Magnetic Resonance Imaging / Q000379:methods*; D008280:Magnetics; D046529:Microscopy, Electron, Transmission; D015394:Molecular Structure; D053758:Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D010316:Particle Size; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011073:Polyamines / Q000737:chemistry; D011095:Polyethylenes / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D014162:Transfection / Q000379:methods
chemical_list: D028581:Lanthanoid Series Elements; D011073:Polyamines; D011095:Polyethylenes; D011108:Polymers; D005063:Europium; D004247:DNA; D005682:Gadolinium; D016650:Fluorescein-5-isothiocyanate