title: Intracellular trafficking of polyamidoamine-poly(ethylene glycol) block copolymers in DNA delivery.
abstract: The delivery of nucleic acids has the potential to revolutionize medicine by allowing previously untreatable diseases to be clinically addressed. Viral delivery systems have shown immunogenicity and toxicity dangers, but synthetic vectors have lagged in transfection efficiency. Previously, we developed a modular, linear-dendritic block copolymer architecture with high gene transfection efficiency compared to commercial standards. This rationally designed system makes use of a cationic dendritic block to condense the anionic DNA and forms complexes with favorable endosomal escape properties. The linear block provides biocompatibility and protection from serum proteins, and can be functionalized with a targeting ligand. In this work, we quantitate performance of this system with respect to intracellular barriers to gene delivery using both high-throughput and traditional approaches. An image-based, high-throughput assay for endosomal escape is described and applied to the block copolymer system. Nuclear entry is demonstrated to be the most significant barrier to more efficient delivery and will be addressed in future versions of the system.
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Daniel K
authors.initials: DK
authors.lastname: Bonner
authors.forename: Cheuk
authors.initials: C
authors.lastname: Leung
authors.forename: Jane
authors.initials: J
authors.lastname: Chen-Liang
authors.forename: Loice
authors.initials: L
authors.lastname: Chingozha
authors.forename: Robert
authors.initials: R
authors.lastname: Langer
authors.forename: Paula T
authors.initials: PT
authors.lastname: Hammond
mesh_terms: D021581:Active Transport, Cell Nucleus; D001672:Biocompatible Materials; D001692:Biological Transport; D004247:DNA / Q000008:administration & dosage*; D011992:Endosomes / Q000378:metabolism*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D057166:High-Throughput Screening Assays; D006801:Humans; D011073:Polyamines / Q000493:pharmacokinetics* / Q000627:therapeutic use; D011092:Polyethylene Glycols / Q000493:pharmacokinetics* / Q000627:therapeutic use; D011108:Polymers / Q000493:pharmacokinetics / Q000627:therapeutic use
chemical_list: D001672:Biocompatible Materials; C531249:Poly(amidoamine); D011073:Polyamines; D011108:Polymers; D011092:Polyethylene Glycols; D004247:DNA