TI:Investigation on activities of hammerhead ribozyme embedded in genomic RNA of hepatitis delta virus
AU:LIXiaojuan, KUANG Ersheng , DAI Wei , YANG Fuhua , WANG Min , WANG Huosheng , ZHOU Boping
AD:Municipal Institute of Hepatology , Shenzhen Eastlake Hospital , Shenzhen 518020 ,China
【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 vivo1 Methods 　In vitro activity of ribozyme or HDV2driven ribozyme was assessed by incubating with the [α232 P]2ATP labeled HBV RNA substrates at different temperature1 Huh27 cells were cotransfected with ribozyme or HDV2 ribozyme chimera and HBV genome , by which inhibition of ribozymes on HBV transcription in vivo were examined1 Results 　The results indicated that both temperature and secondary structure influenced the cleavage activity of HDV2driven ribozyme significantly1 When the factors were eliminated , the HDV2driven ribozyme could act as well as its counterpart naked ribozyme1 While in cultured cells the HDV2driven ribozyme had higher inhibition to HBV gene expression than that of ribozyme alone1 Conclusion 　The results demonstrated that HDV may weaken the activity of embedded ribozyme in vitro , but make it enhanced in cultured cells1 Thus , this study could provide a useful evidence to develop HDV as vector for liver2special delivery of ribozyme to against chronic HBV infection1
【Key words】　Hepatitis delta virus ; 　RNA , catalytic ; 　Vehicles ; 　Gene therapy