title: Hammerhead Ribozymes Suppress HBV(adr) in HepG2 Cells.
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.
authors.affiliation: Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai,200031, China. qiguor@sunm.shcnc.ac.cn
authors.forename: Wen-Chao
authors.initials: WC
authors.lastname: Zheng
authors.forename: Chang-De
authors.initials: CD
authors.lastname: Lu
authors.forename: Yu-Ying
authors.initials: YY
authors.lastname: Kong
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wang
authors.forename: Guo-Rong
authors.initials: GR
authors.lastname: Qi