Table 1. Mutant template telomerase RNA (MT-mTer) reprograms telomerase enzyme to add mutated telomeric repeats.
When MT-mTer and MT-mTer/siRNA are over-expressed, significantly more telomeric products (270% and 350%, respectively) are detected using a TTTGGG-specific (“4A10A”) reverse primer in the second (amplification) step of RT-PCR TRAP, an effect which is preserved even when just dGTP and dTTP are added in the first (extension) step of the reaction. Activity values are % of vector control and are means of triplicates; all values differ from vector control with statistical significance (p<0.01).
Telomerase activity (as % of vector control)* | ||||
---|---|---|---|---|
** Amplification: ACX reverse primer (specific to wild-type TTAGGG) | ** Amplification: 4A10A reverse primer (specific to mutant TTTGGG) | |||
** Extension: 4 dNTPs | ** Extension: dGTP & dTTP | ** Extension: 4 dNTPs | ** Extension: dGTP & dTTP | |
Vector Control | 100 | 0 | 0 | 5 |
MT-mTer | 100 | 0 | 270 | 270 |
MT-mTer/siRNA | 5 | 0 | 350 | 350 |
All activity values are means of triplicates
Extension and amplification steps illustrated in Figure 2D