Development of a chimeric DNA-RNA hammerhead ribozyme targeting SARS virus.
OBJECTIVE
Severe acute respiratory syndrome (SARS) is a severe pulmonary infectious disease caused by a novel coronavirus. To develop an effective and specific medicine targeting the SARS-coronavirus (CoV), a chimeric DNA-RNA hammerhead ribozyme was designed and synthesized using a sequence homologous with the mouse hepatitis virus (MHV).


METHODS
Chimeric DNA-RNA hammerhead ribozyme targeting MHV and SARS-CoV were designed and synthesized.To confirm its activity, in vitro cleavage reactions were performed with the synthesized ribozyme. Effects of the chimeric ribozyme were evaluated on multiplication of MHV. Effects of the chimeric ribozyme on expression of SARS-CoV were evaluated in cultured 3T3 cells.


RESULTS
The synthetic ribozyme cleaved the synthetic target MHV and SARS-CoV RNA into fragments of predicted length. The chimeric DNA-RNA hammerhead ribozyme targeting SARS-CoV significantly inhibited multiplication of MHV in DBT cells by about 60%. The chimeric DNA-RNA hammerhead ribozyme targeting SARS-CoV significantly inhibited the expression of SARS-CoV RNA in 3T3 cells transfected with the recombinant plasmid. The chimeric DNA-RNA ribozyme targeting SARS-CoV significantly inhibited MHV viral activity and expression of recombinant SARS RNA in vitro.


CONCLUSIONS
These findings indicate that the synthetic chimeric DNA-RNA ribozyme could provide a feasible treatment for SARS.
Fukushima
Akiko
A
Division of Gastroenterology and Hepatology, Department of Medicine, Nihon University School of Medicine, Tokyo, Japan.
Fukuda
Noboru
N
Lai
Yimu
Y
Ueno
Takahiro
T
Moriyama
Mitsuhiko
M
Taguchi
Fumihiro
F
Iguchi
Akifumi
A
Shimizu
Kazushi
K
Kuroda
Kazumichi
K
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D002460:Cell Line; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D020871:RNA Stability; D016337:RNA, Catalytic / Q000235:genetics / Q000494:pharmacology*; D012367:RNA, Viral / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D016337:RNA, Catalytic; D012367:RNA, Viral; C427204:hammerhead ribozyme