Antiviral
Activity of a Hammerhead Ribozyme against HBV in HepG2.2.15 Cells
FENG Ying, KONG Yu-Ying, WANG Yuan*,
QI Guo-Rong*
( Institute of Biochemistry and Cell Biology, Shanghai Institutes for
Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031,
China )
Abstract The carboxy
terminal arginine rich domain of core protein is highly conserved among HBV
subtypes, and plays important
roles in the packaging of pregenomic RNA and viral replication. Therefore, the
3＇
highly conserved region of HBV C gene is an attractive target for antiviral
therapy mediated by ribozymes. A hammerhead ribozyme RzC, targeting the above
site, was designed; meanwhile, as
controls, the disabled form of RzC, dRzC, was also designed. In order to
investigate its antiviral effects in cultured cells, in vitro-transcribed
ribozymeRzC, dRzC and ribozyme expression vectors pCRzC, pCdRzC were
delivered, respectively, into
HepG2.2.15 cells ( clonal cells derived from HepG2 cells that contain
integrated HBV ayw DNA). The preliminary results demonstrated that in
vitro-transcribed ribozyme RzC had weak inhibition on HBV replication,
possibly due to its quick degradation by RNases in cells, while the
endogenously expressed ribozyme RzC showed significant inhibitory effect on HBV
replication. In conclusion, the results suggested the possibility of the
hammerhead ribozyme RzC to be used for the gene therapy of HBV infection.
Key words hammerhead ribozyme; HBV; HepG2.2.15 cells
*Corresponding authors: QI
Guo-Rong：
Tel, 86-21-64374430-5230; Fax, 86-21-64338357; e-mail, [email protected]
；
WANG Yuan： Tel, 86-21-64374430-5326; Fax,
86-21-64338357; e-mail, wan[email protected]