In trypanosomatids, the mechanisms of gene expression regulation are not yet well understood. The genes are organized into long polycistronic transcription units separated by intergenic regions that may contain the signaling information for nucleic acid processing. Poly-dinucleotides are frequent in these regions and have been proposed to be involved in gene expression regulation. We analyzed their frequency in CDS-flanking sequences of sense strands in Trypanosoma cruzi and established that all but poly[dC-dC], poly[dC-dG], and poly[dG-dG] are significantly more frequent than expected by chance. Poly[dT-dG](.)[dC-dA] is among the longest and most frequent polydinucleotides and shows a remarkable strand asymmetry. Furthermore, electrophoretic mobility shift assays using T. cruzi epimastigotes nuclear extracts demonstrated the existence of at least, one sequence specific single-strand binding activity for each strand. These results strongly suggest that poly[dT-dG](.)[dC-dA] sequence is involved in regulatory mechanisms of relevance for the parasite biology.