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  • Title: [Investigation on activities of hammerhead ribozyme embedded in genomic RNA of hepatitis delta virus].
    Author: Li XJ, Kuang ES, Dai W, Yang FH, Wang M, Wang HS, Zhou BP.
    Journal: Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi; 2005 Mar; 19(1):12-5. PubMed ID: 16201462.
    Abstract:
    OBJECTIVE: To develop HDV as a vehicle to deliver hammerhead ribozyme into hepatocytes, the effects of modified HDV was assessed on the activity of embedded hammerhead ribozyme in vitro and in vivo. METHODS: In vitro activity of ribozyme or HDV-driven ribozyme was assessed by incubating with the [alpha-32 P]-ATP labeled HBV RNA substrates at different temperature. Huh-7 cells were cotransfected with ribozyme or HDV-ribozyme chimera and HBV genome, by which inhibition of ribozymes on HBV transcription in vivo were examined. RESULTS: The results indicated that both temperature and secondary structure influenced the cleavage activity of HDV-driven ribozyme significantly. When the factors were eliminated, the HDV-driven ribozyme could act as well as its counterpart naked ribozyme. While in cultured cells the HDV-driven ribozyme had higher inhibition to HBV gene expression than that of ribozyme alone. CONCLUSION: The results demonstrated that HDV may weaken the activity of embedded ribozyme in vitro, but make it enhanced in cultured cells. Thus, this study could provide a useful evidence to develop HDV as vector for liver-special delivery of ribozyme to against chronic HBV infection.
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