Investigation on activities of hammerhead ribozyme embedded in genomic RNA of hepatitis delta virus.
- Author:
Xiao-juan LI
1
;
Er-sheng KUANG
;
Wei DAI
;
Fu-hua YANG
;
Min WANG
;
Huo-sheng WANG
;
Bo-Ping ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Cell Line, Tumor; Genome, Viral; Hepatitis Delta Virus; enzymology; genetics; Humans; Molecular Sequence Data; Plasmids; genetics; Protein Structure, Secondary; RNA, Catalytic; chemistry; genetics; metabolism; RNA, Viral; genetics; metabolism; Structure-Activity Relationship; Substrate Specificity; Temperature; Transfection
- From: Chinese Journal of Experimental and Clinical Virology 2005;19(1):12-15
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSIn 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.
RESULTSThe 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.
CONCLUSIONThe 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.
