Selective recognition of a C-G base-pair within the parallel DNA triple-helical binding motif was achieved by a third strand containing the base 5-methyl pyrimidin-2-one. The third strand affinities (K-D) for a representative 15-mer duplex sequence containing all four Watson-Crick base pairs (X-Y) in the center are C-G (26 nM) >> A-T (270 nM) similar to T-A (350 nM) > G-C (ca. 700 nM). (C) 1999 Elsevier Science Ltd. All rights reserved.