Characterization of interpolyelectrolyte complexes between double-stranded DNA and polylysine comb-type copolymers having hydrophilic side chains.
The polyionic interaction between DNA and polycations grafted with hydrophilic dextran side chains was evaluated. The comb-type copolymers, poly(L-lysine)-graft-dextran, were successfully prepared by employing a reductive amination reaction between epsilon-amino groups of poly(L-lysine) (PLL) and the reductive ends of dextran (Dex). A coupling efficacy on the order of 70% was obtained regardless of intrinsic philicities of the solvents used, either aqueous buffer or DMSO. The resulting graft copolymers, which varied in the degree of grafting and the length of hydrophilic side chains, formed a soluble complex with DNA. They also affected the melting behavior of double-stranded DNA (dsDNA) in different ways. Copolymers having a high degree of grafting thermally stabilized dsDNA without affecting its reversible transition between single-stranded and double-stranded forms. However, copolymers with a low degree of grafting or with a high degree of grafting of short dextran chains impeded the reversibility of this transition. Furthermore, highly grafted copolymers also accelerated the hybridization of DNA strands in a low-ionic strength medium. It is of particular note that these copolymers scarcely altered circular dichroismic signals of dsDNA even when the copolymers were added in excess. This suggested that the copolymer interacted with dsDNA without affecting its native structure or physicochemical properties. Finally, the copolymer even formed a stable complex with a short oligonucleotide (20 bases). We, therefore, concluded that, by regulating the degree of grafting and the molecular weight of grafted side chains, it would be possible to design novel different graft copolymers capable of acting as carriers of functional genes to target cells or tissue.
Maruyama
A
A
Department of Biomolecular Engineering, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori, Yokohama, Japan. amaruyam@bio.titech.ac.jp
Watanabe
H
H
Ferdous
A
A
Katoh
M
M
Ishihara
T
T
Akaike
T
T
D000818:Animals; D002412:Cations; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry*; D003911:Dextrans / Q000737:chemistry*; D004121:Dimethyl Sulfoxide; D004355:Drug Stability; D004563:Electrochemistry; D004587:Electrophoresis, Agar Gel; D006358:Hot Temperature; D008970:Molecular Weight; D009391:Nephelometry and Turbidimetry; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D009994:Osmolar Concentration; D011067:Poly dA-dT / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry*; D012995:Solubility; D014867:Water
D002412:Cations; D003911:Dextrans; D014867:Water; D011107:Polylysine; D011067:Poly dA-dT; D004247:DNA; D004121:Dimethyl Sulfoxide