Abstract:Objective:To investigate the interfering effect of the small hairpin RNA (shRNA) targeted to human telomere retrotranscriptase (hTERT) gene and its silence effect on telomerase hTERT gene expression in HeLa cells.
Methods :After preparation of the shRNA in-vitro, the shRNA was transfected to HeLa cells by the calcium phosphate method. Telomerase activity was detected by TRAP-silver stain and PCR-EIA.
Results:The telomerase activity of HeLa cells decreased significiantly after transfected with the 46-base small hairpin RNA.
Conclusions:shRNA (targeted to human telomerase hTERT gene) prepared by this method displays a significant silence effect on telomerase gene expression. It could be a new approach of obtaining shRNA for RNAi research in tumor inhibition.