Synthesis of ribozyme against vascular endothelial growth factor_(165) and its biological activity in vitro

摘要

AIM: To investigate the designation, synthesis and biological activity of against vascular endothelial growth factor_(165) (VEGF_(165)) ribozyme. METHODS: The ribozyme against VEGF_(165) was designed with computer. The transcriptional vector was constructed which included the anti-VEGF_(165) ribozyme and 5′, 3′ self-splicing ribozymes. The hammerhead ribozyme and substrate VEGF_(165) 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-VEGF_(165) 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%. CONCLUSION: The anti-VEGF_(165) ribozyme designed with computer can cleave VEGF_(165) mRNA effectively.