3.2. Attenuation of telomerase activity by shRNA against hTERT
The observed reduction of hTERT transcript level should reflect in its
functional activity. To assess shRNA efficiency in telomerase inhibition,
extracts from transfected constructs were analyzed with Telomeric Repeat
Amplification Protocol (TRAP) assay with serially diluted (100 ng
to 1000 ng) cellular protein. The TRAP assay with 500 ng protein
showed visible reduction in signal intensity of telomerase extended ladders
in HeLa Tet-On cells transfected with shRNA hTERT when compared
to control (Fig. 2E) showing that shRNA against hTERT indeed
reduces the telomerase activity in HeLa cell line.