The 5'-end of hTERT mRNA is a good target for hammerhead ribozyme to suppress telomerase activity.
Because the expression level of hTERT, a catalytic subunit of human telomerase, is a rate-limiting determinant of telomerase activity, hTERT mRNA would be an excellent target of hammerhead ribozymes for the regulation of telomerase activity. We studied the efficiency of several hammerhead ribozymes targeting hTERT mRNA by transient and stable transfection procedures. To screen the potency of the ribozymes, transient ribozyme transfection and telomerase determination were performed. The ribozyme targeting 13 nucleotides downstream from the 5'-end of hTERT mRNA (13-ribozyme) exhibited the strongest telomerase-inhibitory activity, and the ribozyme to target 59 nucleotides upstream from the poly(A) tail showed clear activity. A stable transfection study confirmed that the 13-ribozyme suppressed telomerase. These observations suggest that the 13-ribozyme can regulate telomerase activity and may possess potential for cancer therapy.
Yokoyama
Y
Y
Department of Obstetrics and Gynecology, Gifu University School of Medicine, 40 Tsukasa-machi, Gifu, 500-8705, Japan. yokoyama@cc.gifu-u.ac.jp
Takahashi
Y
Y
Shinohara
A
A
Wan
X
X
Takahashi
S
S
Niwa
K
K
Tamaya
T
T
D020121:5' Untranslated Regions / Q000235:genetics / Q000378:metabolism*; D017209:Apoptosis; D001483:Base Sequence; D020134:Catalytic Domain / Q000235:genetics*; D002455:Cell Division; D004268:DNA-Binding Proteins; D016889:Endometrial Neoplasms / Q000201:enzymology / Q000235:genetics / Q000473:pathology / Q000628:therapy; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007700:Kinetics; D009690:Nucleic Acid Conformation; D011061:Poly A / Q000235:genetics / Q000378:metabolism; D012313:RNA; D016372:RNA, Antisense / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000627:therapeutic use; D013379:Substrate Specificity; D019098:Telomerase / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D020121:5' Untranslated Regions; D004268:DNA-Binding Proteins; D016372:RNA, Antisense; D016337:RNA, Catalytic; C422952:telomerase RNA; D011061:Poly A; D012313:RNA; D019098:Telomerase