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Promotion of Duplex and Triplex DNA Formation by Polycation Comb-Type Copolymers

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Triplex DNA has attracted considerable interest recently because of its possible biological functions in vivo and its wide variety of potential applications, such as regulation of gene expression, site-specific cleavage of duplex DNA, mapping of genomic DNA, and gene-targeted mutagenesis (1 3) . A triplex is usually formed through the sequence-specific interaction of a single-stranded homopyrimidine or homopurine triplex-forming oligonucleotide (TFO) with the major groove of the homopurine-homopyrimidine stretch in duplex DNA (1 5 ). In the pyrimidine motif triplex, a homopyrimidine TFO binds parallel to the homopurine strand of the target duplex by Hoogsteen hydrogen bonding to form T●A:T and C+● G:C triplets (● and : represent Hoogsteen hydrogen bonding and Watson Crick base pairing, respectively). (1 5 ). Because the cytosine bases in a homopyrimidine TFO are to be protonated to bind with the guanine bases of the G:C duplex, the formation of the pyrimidine motif triplex needs an acidic pH condition, and is thus unstable at physiological pH (6 8 ). On the other hand, in the purine motif triplex, a homopurine TFO binds antiparallel to the homopurine strand of the target duplex by reverse Hoogsteen hydrogen bonding to form A●A:T (or T●A:T) and G●G:C triplets (1 5 ). Although the purine motif triplex is pH-independent, triplexes involving guanine-rich TFOs are inhibited by physiological concentrations of certain monovalent cations (M+) , especially K+ (9 ,10 )
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