Development of a novel gene delivery scaffold utilizing colloidal gold–polyethylenimine conjugates for DNA condensation

Abstract

e have developed a novel gene delivery scaffold based on DNA plasmid condensation with colloidal gold/polyethylenimine conjugates. This scaffold system was designed to enable systematic study of the relationships between DNA complex physical properties and transfection efficiency. Using an enhanced green fluorescent protein-coding reporter plasmid and a Chinese hamster ovary cell line, we have measured the transfection efficiencies of our complexes using flow cytometry and their cytotoxicities using the trypan blue assay. We have also assayed complex particle morphologies using atomic force microscopy, photon correlation spectroscopy, and a novel plasmon absorbance peak position analysis. We achieved comparable rates of transfection relative to the commonly used polycationic condensation agents calcium phosphate and LipofectAMINEt, with comparably low cytotoxicities. In addition, by manipulating colloidal gold concentration, we could partially decouple complex physical properties including charge ratio, size, DNA loading, and polyethylenimine concentration. Our morphological analyses showed that complexes with a diameter of a few hundred nanometers and a charge ratio of B8 perform best in our transfection efficiency assays. The use of colloidal gold as a component in our delivery system provides a versatile system for manipulating complex properties and morphology as well as a convenient scaffold for planned ligand conjugation studies.

Biocompatibility of Fe3O4@Au composite magnetic nanoparticles in vitro and in vivo

“…32 Similarly, the protective properties of the gold coating have also been reported by others. 33 However, the literature also reported conflicting data regarding the cytotoxicity of gold nanoparticles in vitro. For example, some researchers demonstrated that different sized gold nanoparticles with varying surface chemistries were not toxic to human cells.…”

In vivo expression of myosin essential light chain using plasmid expression vectors in regenerating frog skeletal muscle

“…Recently, encouraging results have been obtained that demonstrate transfection efficiency of plasmid expression vectors are greatly enhanced when the pDNA is complexed to various biopolymers. [11][12][13][14] Studies are ongoing in our laboratory to test the efficacy of various biopolymer-pDNA complexes to produce higher and more consistent levels of trans-myosin expression following in vivo gene transfer.…”

“…The AT muscle was pulverized in liquid N 2 , and total RNA was isolated using the TRIzol reagent (Invitrogen, Carlsbad, CA, USA). Total RNA was reverse transcribed (Superscript II, Invitrogen) to cDNA using an oligo (dT) [12][13][14][15][16][17][18] primer. Full-length MLC1 f was amplified from the cDNA using a sense primer (5 0 -TTTATTGG GGCGGCCGCCTAGGCACTGGGCTGAGGAT-3 0 ) and -TTTGGTGGTACCGCCCTAAACTTT GGACAGAT-3 0 ) complimentary to part of the 5 0 UTR and 3 0 UTR, respectively (NotI and KpnI sites underlined).…”

“…Recent successes in enhanced plasmid gene delivery using nanopolymer agents have rekindled the focus on plasmids as effective gene carriers for therapeutic purposes. [11][12][13][14] Intensive efforts have been undertaken to understand the molecular basis of skeletal muscle myosin structure and function, 15,16 including the relationship between mutations and disease. 17,18 Frog muscle provides an exceptional opportunity to exploit gene transfer to study the structural basis of myosin function.…”

Rheological Parameters Assessment in Serum, Plasma and Whole Blood of Rats after Administration of Gold Nanoparticles of Different Sizes: In vivo

“…GNPs are used as immunostaining marker particles for electron microscopy and as chromophores for immunoreactions and nucleic acid hybridisation [12,13]. Their application for gene delivery into cells has also been reported [14][15][16][17]. In addition, GNPs have attracted attention as photothermal agents in hyperthermia treatment [18].…”