title: Multifunctional non-viral gene vectors with enhanced stability, improved cellular and nuclear uptake capability, and increased transfection efficiency.
abstract: We have developed a new multifunctional, non-viral gene delivery platform consisting of cationic poly(amine-co-ester) (PPMS) for DNA condensation, PEG shell for nanoparticle stabilization, poly(γ-glutamic acid) (γ-PGA) and mTAT (a cell-penetrating peptide) for accelerated cellular uptake, and a nuclear localization signal peptide (NLS) for enhanced intracellular transport of DNA to the nucleus. In vitro study showed that coating of the binary PPMS/DNA polyplex with γ-PGA promotes cellular uptake of the polyplex particles, particularly by γ-glutamyl transpeptidase (GGT)-positive cells through the GGT-mediated endocytosis pathway. Conjugating PEG to the γ-PGA led to the formation of a ternary PPMS/DNA/PGA-g-PEG polyplex with decreased positive charges on the surface of the polyplex particles and substantially higher stability in serum-containing aqueous medium. The cellular uptake rate was further improved by incorporating mTAT into the ternary polyplex system. Addition of the NLS peptide was designed to facilitate intracellular delivery of the plasmid to the nucleus--a rate-limiting step in the gene transfection process. As a result, compared with the binary PPMS/LucDNA polyplex, the new mTAT-quaternary PPMS/LucDNA/NLS/PGA-g-PEG-mTAT system exhibited reduced cytotoxicity, remarkably faster cellular uptake rate, and enhanced transport of DNA to the nucleus. All these advantageous functionalities contribute to the remarkable gene transfection efficiency of the mTAT-quaternary polyplex both in vitro and in vivo, which exceeds that of the binary polyplex and commercial Lipofectamine™ 2000/DNA lipoplex. The multifunctional mTAT-quaternary polyplex system with improved efficiency and reduced cytotoxicity represents a new type of promising non-viral vectors for the delivery of therapeutic genes to treat tumors.
authors.affiliation: Department of Biomedical Engineering, School of Engineering, Sun Yat-sen University, Guangzhou, Guangdong 510006, China. bmeliu@gmail.com.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Yang
authors.forename: Zhaozhong
authors.initials: Z
authors.lastname: Jiang
authors.forename: Zhong
authors.initials: Z
authors.lastname: Cao
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.forename: Di
authors.initials: D
authors.lastname: Gao
authors.forename: Xingen
authors.initials: X
authors.lastname: Luo
authors.forename: Xiaofang
authors.initials: X
authors.lastname: Zhang
authors.forename: Huiyan
authors.initials: H
authors.lastname: Luo
authors.forename: Qing
authors.initials: Q
authors.lastname: Jiang
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
mesh_terms: D002467:Cell Nucleus / Q000737:chemistry* / Q000235:genetics*; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D004355:Drug Stability; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D053769:Nanocapsules / Q000008:administration & dosage / Q000737:chemistry* / Q000648:ultrastructure; D058726:Nanoconjugates / Q000008:administration & dosage / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D011108:Polymers / Q000737:chemistry; D014162:Transfection / Q000379:methods*
chemical_list: D053769:Nanocapsules; D058726:Nanoconjugates; D011108:Polymers; D004247:DNA