Several mechanisms have been proposed to explain berries' anticancer effects. However, no previously published studies have investigated berries' anti-telomerase activity. In this study, the anti-telomerase activity of blackberry crude extract was analyzed in six human colorectal cancer (CRC) cell lines by TRAP assay. The peripheral blood mononuclear cells (PBMCs) from a healthy donor were used as a normal control. We also examined the effect of blackberry on the human telomerase RNA (hTR) mRNA level and on human telomerase reverse transcriptase (hTERT) expression and promoter methylation in CRC cells. Blackberry extract significantly inhibited the growth of six CRC cell lines in a dose-dependent manner. Telomerase activity of CRC cells incubated with the IC50 concentration of berry's extract for 48 and 72 h decreased by 15%-37.5% and 43.23%-62.5% (P < 0.05), respectively. In cell-free assays, treatment with as little as 7 µl/ml of berry juice completely blocked telomerase activity in CRC cell lysates. Berry was much less effective in inhibiting telomerase activity in normal PBMCs than CRC cells. Berry treatment reduced hTERT expression and its promoter methylation in CRC cell lines, but the expression of hTR was less influenced by the treatment. Our data indicate that telomerase inhibition is a key mechanism by which blackberry exerts its anticancer effects in CRC cells.