Synthesis of ribozyme against vascular endothelial growth factor165 and its biological activity in vitro.
OBJECTIVE
To investigate the designation, synthesis and biological activity of against vascular endothelial growth factor165 (VEGF165) ribozyme.


METHODS
The ribozyme against VEGF165 was designed with computer. The transcriptional vector was constructed which included the anti-VEGF165 ribozyme and 5', 3' self-splicing ribozymes. The hammerhead ribozyme and substrate VEGF165 mRNA were synthesized through transcription in vitro. The cleavage activity of the ribozyme on target RNA was observed in a cell-free system.


RESULTS
The anti-VEGF165 ribozyme was released properly from the transcription of pGEMRz212 cleaved by 5' and 3' self-splicing ribozymes which retained its catalytic activity, and the cleavage efficiency of ribozyme reached 90.7%.


CONCLUSIONS
The anti-VEGF165 ribozyme designed with computer can cleave VEGF165 mRNA effectively.
Gu
Zhong-Ping
ZP
Department of Thoracic Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, Shaanxi Province, China. zhongpg@xaonline.com.cn
Wang
Yun-Jie
YJ
Wu
Yu
Y
Li
Jin-Ge
JG
Chen
Nong-An
NA
D003198:Computer Simulation; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000138:chemical synthesis* / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism*
D016337:RNA, Catalytic; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A