Transfection poly

Uchegbu and coworkers have studied the complexation and delivery of DNA using a unique poly(amino acid)-based polymer vesicle. A polymer of either poly (L-lysine) or poly(L-omithine) was functionalized with methoxy-poly(ethylene glycol) (mPEG) and hydrophobic palmitic acid chains to synthesize an amphiphilic triblock of either mPEG-6-poly(L-lysine)-6-palmitoyl or mPEG-Z>-poly(L-omithine)-6-palmitoyl. Vesicles formed from these polymers were complexed with DNA and showed improved transfection in vitro over poly(amino acid) complexed with DNA or DNA alone [82]. 

The advantage of mRNA over plasmid transfection is the ability of in vitro transcription to allow precise control over features contained within the mRNA (Humphreys et al., 2005 Pillai et al., 2005 Westman et al, 2005). For example, mRNA can be prepared either with or without the physiological m7G(5/)ppp(5/)G cap structure and S poly(A) tail, which are important mediators of canonical translation initiation (Gallie, 1991 Hentze et al., 2006 Iizuka et al, 1994 Kahvejian et al, 2005 Tarun and Sachs, 1995). 

Peng SF, Yang MJ, Su CJ et al (2009) Effects of incorporation of poly(y-glutamic acid) in chitosan/DNA complex nanoparticles on cellular uptake and transfection efficiency. Biomaterials 30 1797-1808... 

Poly-L-lysine was one of the original reagents used to condense DNA into a transfecting particle (21) and numerous studies have been widely reported with these reagents. Oligolysine peptides as short as (K)i6 will also package pDNA, alone (polyplex) or in combination with lipids (lipopolyplex) (22-26). In both lipopolyplex and polyplex formulations, the DNA is condensed... 

Another cationic polymer, poly- -(4-aminobutyl)-L-glycolic) acid (PLAGA) has been shown to condense DNA efficiently and also to be less cytotoxic than PLL. PLAGA is biodegradable, not toxic to the cells, and enhances transfection in cultured cells (202). 

Aral, C., Akbuga, J. (2003). Preparation and in vitro transfection efficiency of chitosan microspheres, containing plasmid DNA poly (L-lysine) complexes. J. Pharm. Phantn. Sci., 6(3), 321-326. 

Chemg, J.Y., Van de Wetering, R, Talsma, H., Crommelin, D.J.A., Hennink, W.E. (1996). Effect of size and serum proteins on transfection efficiency of poly ((2-dime-thylamino)ethyl methacrylate)-plasmid nanoparticles. Pharm. Res., 13, 1038-1042. 

Cherng, J.Y., Schuurmans-Nieuwenbroek, N.M., Jiskoot, W., Talsma, H., et al. (1999). Effect of DNA topology on the transfection efficiency of poly((2-dimethy-lamino)ethyl methacrylate)-plasmid complexes. J. Control Release, 60, 343-353. 

J.S. (1999). Transfection and physical properties of various saccharide, poly(ethylene glycol), and antibody-derivatizedpolyethylenimines (PEI). J. Gene Med., 1 (3), 210-222. 

Poly(/V-alk i-4-vinylpyridinium) salts (PVP) Linear, quaternary ammonium salt In vitro transfection in mammalian and bacillus cells (Kabanov etal., 1989)... 

Poly(2- dimethylaminoethyl methacrylate) (PDMAEM) Linear, tertiary amino groups In vitro transfection (Chemg etal., 1996)... 

Poly[alpha-(4-aminobutyl)-L-glycolic acid] (PAGA) Block and graft copolymers Linear, primary amino groups, biodegradable In vitro and in vivo transfections, including therapeutic genes (Kohetal., 2000)... 

Poly(ethylene oxide )-g-poly (/.-lysine) (PEO-g-PLL) Graft copolymer In vitro and in vivo transfection, delivery of oligonucleotides Choi etal, 1998)... 

Polyfection was carried out with pCMVLuc (a 5 kb pDNA) on HepG2 cells for four hours at 37 °C. After 48 hours, the transfection efficiency was determined from Luciferase activity in cells measured by luminescence (RLU) and the cytotoxicity was evaluated by using the colorimetric MTT assay, dp is the poly lysine degree of polymerization. The transfection efficiency is scored on a 0 to 100 scale where 0 and 1 00 correspond to an absence of transfection and the most effective transfection, respectively compared to the pDNA alone. His is histidyl residue. 

---