Anti-viral activity of hairpin ribozyme directed against HBV core region in vitro.
To study the preparation and cleavage of hairpin ribozyme (HpRz) directed against the transcript of HBV core region in vitro, HRz gene designed by computer targeting the transcript of HBV core gene was cloned into the vector p1.5 between 5'-cis-Rz and 3'-cis-Rz. 32P-labeled HpRz transcript proved whether the vector fit for the preparation of hairpin ribozyme. 32P-labeled pKC transcript containing HBV core region as targets-RNA was transcribed by using T7 RNA polymerase and purified by PAGE. Cold HpRz transcript was incubated with 32P-labeled target-RNAs under different conditions and radioautographed after denaturing polyacrylamide gel electrophoresis. The results showed that HpRz had the ability of cleavage at 37 degrees C and 12 mmol/L MgCl2 and the design of ribozyme was correct. It is concluded that HpRz prepared in vitro possesses specific catalytic activity, indicating that it is possible for HpRz to intracellularly inhibit the replication of HBV. It may be developed into a nucleic acid drug in the treatment of hepatitis B in the future.
Lin
J
J
Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030.
Song
Y
Y
Kong
X
X
Xie
N
N
Wu
X
X
Liu
N
N
Wang
N
N
Cao
E
E
Jin
Y
Y
D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D018014:Gene Transfer Techniques; D006512:Hepatitis B Core Antigens / Q000187:drug effects* / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D016337:RNA, Catalytic / Q000494:pharmacology*; D012367:RNA, Viral / Q000235:genetics; D014158:Transcription, Genetic; D014779:Virus Replication / Q000235:genetics
D000998:Antiviral Agents; D006512:Hepatitis B Core Antigens; D016337:RNA, Catalytic; D012367:RNA, Viral; C418481:hairpin ribozyme