Inhibition of telomerase activity by ribozyme targeted to human telomerase transcriptase

• Published in: Sheng wu hua hsüeh yü sheng wu wu li hsüeh pao Vol. 34; no. 3; p. 323
• Main Authors: Qu, Y, Liu, S Q, Peng, W Z, Liu, B L
• Format: Journal Article
• Language: Chinese
• Published: China 01.05.2002
• Subjects: DNA-Binding Proteins; HeLa Cells; Humans; Reverse Transcriptase Polymerase Chain Reaction; RNA, Catalytic - metabolism; RNA, Catalytic - pharmacology; RNA, Messenger - drug effects; RNA, Messenger - metabolism; Telomerase - antagonists & inhibitors; Telomerase - genetics; Telomerase - metabolism; Transfection
• ISSN: 0582-9879

Human telomerase reverse transcriptase (hTERT) is the catalytic subunit and the key factor which controls the telomerase activity,so it is the best choice to inhibit telomerase through controling hTERT expression.In this work,a hammer head ribozyme directed against the hTERT mRNA (hTERTRZ) was designed and synthesized to serve as a telomerase inhibitor. In order to test its in vitro cleavage activity, two in vitro transcription plasmids containing hTERTRZ and hTERT gene respectively were constructed. Ribozyme RNA and DIG-labeled-hTERT were synthesized by in vitro transcription. In vitro cleavage reactions were carried out by mixing the hTERTRZ with DIG-labeled-hTERT under different reaction conditions, and cleavage bands were detected by digoxin chemiluminescent assay. hTERTRZ showed a specific cleavage activity against the hTERT used as template. To investigate its in vivo effect of telomerase inhibition in tumor cells, a eukaryotic expression plasmid containing the hTERT ribozyme gene was introduced into He