Design and synthesis of vascular endothelial growth factor specific multi-site ribozyme and its cleaved activity in vitro

    GU Zhongª²Ping1, WANG Yaoª²Cheng2, ZHOU Yongª²An1, LI Jinª²Ge3, ZHANG Tao1 ,CHEN Nongª²An4

    1Department of Thoracic Surgery, 2Department of Radiology, 3Department of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi¬ðan 710038,China,4Shanghai Institute of Biochemistry, Academia Sinica of China,Shanghai 200020,China

    ¡¾Abstract¡¿ AIM: To design and synthesize a multiª²site ribozyme against VEGF165 and to investigate its cleaved activity in vitro. METHODS: Specific singleª²site (Rz1, Rz2, Rz3) and multiª²site ribozymes (Rz123) against VEGF165 were designed by computer analysis of the secondary structure of the ribozyme and the target RNA flanking sequence around the cleavage sites. The transcriptional vector was constructed, which included the multiª²site ribozyme and 5¡äª², 3¡äª²cis selfª²splicing ribozymes. The cleavage activity of the multiª²site ribozyme on target RNA was observed. RESULTS: The multiª²site ribozyme was released properly from the transcription of the selfª²splicing vector with cleavage by 5¡äª², 3¡äª²cis ribozymes. The multiª²site ribozyme cleaved the target RNA and increased the cleavage efficiency. The cleavage rate was (37¡À10)% (Rz1), (51¡À8)% (Rz2 ), (68¡À15)% (Rz3) and (88¡À11)% (Rz123), respectively. CONCLUSION: The synthesized multiª²site ribozyme designed with computer can effectively cleave target RNA. ......