title: Multivalent dendrimer vectors with DNA intercalation motifs for gene delivery.
abstract: Poly(amido amine) (PAMAM) dendrimers constitute an important class of nonviral, cationic vectors in gene delivery. Here we report on a new concept for dendrimer vector design based on the incorporation of dual binding motifs: DNA intercalation, and receptor recognition for targeted delivery. We prepared a series of dendrimer conjugates derived from a fifth generation (G5) PAMAM dendrimer, each conjugated with multiple folate (FA) or riboflavin (RF) ligands for cell receptor targeting, and with 3,8-diamino-6-phenylphenanthridinium ("DAPP")-derived ligands for anchoring a DNA payload. Polyplexes of each dendrimer with calf thymus dsDNA were made and characterized by surface plasmon resonance (SPR) spectroscopy, dynamic light scattering (DLS) and zeta potential measurement. These studies provided evidence supporting polyplex formation based on the observation of tight DNA-dendrimer adhesion, and changes in particle size and surface charge upon coincubation. Further SPR studies to investigate the adhesion of the polyplex to a model surface immobilized with folate binding protein (FBP), demonstrated that the DNA payload has only a minimal effect on the receptor binding activity of the polyplex: KD = 0.22 nM for G5(FA)(DAPP) versus 0.98 nM for its polyplex. Finally, we performed in vitro transfection assays to determine the efficiency of conjugate mediated delivery of a luciferase-encoding plasmid into the KB cancer cell line and showed that RF-conjugated dendrimers were 1 to 2 orders of magnitude more effective in enhancing luciferase gene transfection than a plasmid only control. In summary, this study serves as a proof of concept for DNA-ligand intercalation as a motif in the design of multivalent dendrimer vectors for targeted gene delivery.
authors.affiliation: Michigan Nanotechnology Institute for Medicine and Biological Sciences and ‡Department of Internal Medicine, University of Michigan Medical School , Ann Arbor, Michigan 48109, United States.
authors.forename: Pamela T
authors.initials: PT
authors.lastname: Wong
authors.forename: Kenny
authors.initials: K
authors.lastname: Tang
authors.forename: Alexa
authors.initials: A
authors.lastname: Coulter
authors.forename: Shengzhuang
authors.initials: S
authors.lastname: Tang
authors.forename: James R
authors.initials: JR
authors.lastname: Baker
authors.forename: Seok Ki
authors.initials: SK
authors.lastname: Choi
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D050091:Dendrimers / Q000008:administration & dosage* / Q000737:chemistry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007624:KB Cells; D059372:Nucleotide Motifs / Q000235:genetics*
chemical_list: D050091:Dendrimers