Activity identification of ribozyme and U1 snRNA chimeric ribozyme against TGFbeta1 in cell-free system and in hepatic stellate cells.
Transforming growth factorbeta1 (TGFbeta1) is known to be intimately involved in many cellular processes. To explore the mechanism of TGFbeta1 in these processes, the non-chimeric hammerhead ribozyme and U1 snRNA chimeric ribozyme against TGFbeta1 were designed to down-regulate TGFbeta1 expression. The activity of non-chimeric ribozyme and U1 snRNA chimeric ribozyme against TGFbeta1 in vitro and in activated hepatic stellate cells (HSCs) was detected. Cleavage reactions of both ribozymes in vitro demonstrated that non-chimeric ribozyme possessed better cleavage activity in vitro than U1 snRNA chimeric ribozyme. The further study showed U1 snRNA chimeric ribozyme inhibited TGFbeta1 expression more efficiently than non-chimeric ribozyme in transfected HSC cells. So it indicates that the U1 snRNA chimeric ribozyme provides an alternative approach for the research on the precise mechanism of TGFbeta1 in many cellular processes and a potential therapeutic candidate for TGFbeta1-related diseases.
Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Yuhu
Y
Song
Fang
F
Liu
Dean
D
Tian
Xiulan
X
Xue
Nanzhi
N
Liu
Xiaoli
X
Wu
Jusheng
J
Lin
Youxin
Y
Jin
D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002474:Cell-Free System; D017931:DNA Primers; D008099:Liver / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000235:genetics*
D017931:DNA Primers; D016337:RNA, Catalytic; D012342:RNA, Small Nuclear; D016212:Transforming Growth Factor beta; C405483:U1 small nuclear RNA