Design of polymer materials enhancing nucleotide hybridization for anti-gene technology

Abstract

Stabilization of nucleotide hybridization is considered important for improving gene therapy using oligonucleotides. We have designed comb-type copolymer consisting of polycation backbone (polylysine) and hydrophilic side chains as a stabilizer for double and triple helical DNAs. The copolymer considerably increased the thermal stability of triple helical structure but did not affect the reversible transition between triple helical and single-stranded DNA. An in vitro electrophoretic mobility shift assay revealed that the copolymer remarkably increased association constants of both Hoogsteen and reverse Hoogsteen-type triple helix formation. Moreover the triple helix-stabilizing efficiency of the copolymer was significantly higher than that of other oligocations like spermine and spermidine. Not only being good DNA triple helix stabilizer, it has also been shown to accelerate DNA strand exchange reactions between double helical DNA and its complementary oligonucleotides. From these, we conclude that this copolymer is capable of either ‘stabilizing’ or ‘activating’ DNA hybrids, and may useful for gene targeting employing oligonucleotides.