Selective regression of cells expressing mouse cytoskeleton-associated protein 2 transcript by trans-splicing ribozyme.
Cytoskeleton-associated protein 2 (CKAP2) is known to be highly expressed in primary human cancers as well as most cancer cell lines. CKAP2 functions as microtubule stabilizer and probably as cell proliferation inducer, indicating that CKAP2 might be a potential anticancer target. In this study, we developed a specific ribozyme that can replace mouse CKAP2 (mCKAP2) RNA with new transcripts through trans-splicing reaction. This specific RNA replacement resulted in triggering of transgene activity selectively in mammalian cells that express the mCKAP2 RNA. Simultaneously, the ribozyme reduced the expression level of the target RNA in the cells. Noticeably, the ribozyme selectively induced activity of the suicide gene herpes simplex virus thymidine kinase in cells expressing the mCKAP2 RNA and thereby specifically retarded the survival of these cells with ganciclovir treatment. This mCKAP2-specific ribozyme will be useful for validation of the RNA replacement as cancer gene therapy approach in mouse model with syngeneic tumors.
Kim
Areum
A
Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University, Seoul 140-714, Korea.
Ban
Guyee
G
Song
Min-Sun
MS
Bae
Chang-Dae
CD
Park
Joobae
J
Lee
Seong-Wook
SW
D000818:Animals; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000235:genetics*; D015316:Genetic Therapy; D006801:Humans; D051379:Mice; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D020040:Trans-Splicing; D014158:Transcription, Genetic; D019076:Transgenes
C494575:CKAP2 protein, mouse; D003598:Cytoskeletal Proteins; D016337:RNA, Catalytic; D012333:RNA, Messenger