Journal article
Ribozyme cleaves rex/tax mRNA and inhibits bovine leukemia virus expression
Proceedings of the National Academy of Sciences - PNAS, Vol.90(23), pp.10932-10936
12/01/1993
Handle:
https://hdl.handle.net/2376/108530
PMCID: PMC47895
PMID: 7504287
Abstract
Bovine leukemia virus (BLV) encodes at least two regulatory proteins, Rex and Tax. Tax, the transactivating protein, stimulates the long terminal repeat to promote viral transcription and may be involved in tumorigenesis. Rex is involved in the transition from early expression of regulatory proteins to later expression of viral structural proteins. We have targeted ribozymes against the mRNA encoding Rex and Tax. The ribozymes consist of the hammer-head catalytic motif flanked by antisense sequences that hybridize with the complementary rex/tax mRNA. To evaluate cleavage in a cell-free system, we transcribed portions of rex/tax mRNA and incubated them with synthetic RNA ribozymes. A ribozyme was identified that cleaves > 80% of the target RNA. Synthetic DNA encoding this ribozyme was cloned into the expression vector pRc/RSV and transfected into BLV-infected bat lung cells. Intracellular cleavage of rex/tax mRNA was confirmed by reverse transcriptase PCR. In cells expressing the ribozyme, viral expression was markedly inhibited. Expression of the BLV core protein p24 was inhibited by 61%, and reverse transcriptase activity in supernatant was inhibited by 92%. Ribozyme inhibition of BLV expression suggests that cattle expressing these sequences may be able to control BLV replication.
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Details
- Title
- Ribozyme cleaves rex/tax mRNA and inhibits bovine leukemia virus expression
- Creators
- G H Cantor - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040T F McElwain - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040T A Birkebak - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040G H Palmer - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040
- Publication Details
- Proceedings of the National Academy of Sciences - PNAS, Vol.90(23), pp.10932-10936
- Academic Unit
- Paul G. Allen School for Global Animal Health
- Identifiers
- 99900547254001842
- Language
- English
- Resource Type
- Journal article