7,8-Dihydro-8-oxoadenine as a replacement for cytosine in the third strand of triple helices. Triplex formation without hypochromicity.

Oligonucleotides containing thymine and cytosine (or 5-methylcytosine) bases are known to bind to specific homopurine sequences in double-stranded DNA by means of T.AT and C+.GC base triplets. Cytosine in the third strand of such triple helices can be completely replaced by 7,8-dihydro-8-oxoadenine a base which should not require protonation to form base triplets. Experiments using native PAGE and inhibition of triplex-directed photo-cross-linking demonstrate that triplexes with 7,8-dihydro-8-oxoadenine in the third strand are as stable at pH 6.0 as triplexes with 5-methylcytosine. The stability of triplexes with 7,8-dihydro-8-oxoadenine, unlike those with 5-methylcytosine, is not substantially diminished upon raising the pH to 7.4. Surprisingly, triplex formation with an oligonucleotide containing only thymine and 7,8-dihydro-8-oxoadenine was not associated with significant hypochromicity and could not be detected in conventional thermal denaturation experiments.