Hammerhead Ribozymes
Suppress HBV(adr) in HepG2 Cells
ZHENG Wen-Chao,LU Chang-De,KONG Yu-Ying,WANG Yuan,QI
Guo-Rong*
( Shanghai Institute of Biochemistry, the Chinese Academy of Sciences,
Shanghai,200031 )
Abstract Three
hammerhead ribozymes (RS3,RC2 and RC1) targeting to the HBV genome have been
designed. Plasmids were constructed by inserting the genes of naked and
tRNA-embedded ribozymes into RNA trimming vector pRG523 and then were
transferred to eukaryotic expression vector.By the similar cloning method the
shotgun-type plasmids carrying homogeneous RS3 or RtS3 unitconnected in tandem
were obtained. After co-transfecting the above plasmids and HBV genome
containing plasmid into human hepatoma cell line HepG2 respectively and
selection by G418, the HBV-inhibiting activity of different kinds of ribozyme
in G418-resistant cells was achieved by measuring the decrease of HBV-RNA,
progeny DNA and the antigens expressed. The results showed that all the
ribozymes were active with more than 70% inhibition activity against the HBV
and that tRNA-embedded ribozymes had higher activity than naked ribozymes. It
is worth particular interest that shotgun-type ribozymes with the connected
unit in tandem with 8 and 12 units constructed in the plasmid revealed the
highest activity, reaching >90% inhibition.
Key words hammerhead
ribozyme；HBV；HepG2 cells
*Corresponding author：Tel,86-21-64374430；Fax,86-21-64338357；e-mail,[email protected]