骨肉瘤中DNAzymes针对ezrin mRNA作用靶点筛选及体外验证  

Designation and screening of ezrin mRNA target sites for DNAzymes in osteosarcoma and the validation in vitro

作　　者：张国如[1] 王晋[2] 王姚斐[2] 陈运崇[1] 杜秀藩[1] 

机构地区：[1]海南省第三人民医院骨科,海南三亚572000 [2]中山大学附属第一医院骨科,广东广州510080

出　　处：《海南医学》2013年第20期2965-2967,共3页Hainan Medical Journal

基　　金：海南省自然科学基金(编号:310182)

摘　　要：目的对Ezrin mRNA的4个DNAzymes靶点AU1655、AU1751、AU1766、AU1789通过体外实验进行验证,寻找最佳的作用位点。方法同时使用RNAdraw、RNAstructure和OligoWalk 3个程序对Ezrin mRNA的DNAzymes靶点进行了设计和筛选,对最终选出的4个位点AU1655、AU1751、AU1766、AU1789进行实验研究。结果 4个DNAzymes分别命名为Dz1655、Dz1751、Dz1766和Dz1789,酶切反应结果只有Dz1751在预期的位点成功切割了ezrin全长的mRNA,得到了128 bp和1 633 bp两个片段,其他3个DNAzymes均未能有效地完成切割;反应后所剩留底物RNA中位点AU1751最少,位点AU1751是DNAzymes切割ezrin mRNA的最佳位点。结论核酸二级结构联合热动力学参数可以最大程度预测和设计DNAzymes针对ezrin mRNA的作用位点。Objective To verify the ezrin mRNA target sites （AU1655, AU1751, AU1766, and AU1789） for DNAzymes through experiments in vitro. Methods Three programs （RNAdraw, RNAstructure and OligoWalk） were applied to design and screen the ezrin mRNA target sites for DNAzymes, and finally 4 targeting sites （AU1655, AU1751, AU1766, and AU1789） were selected for study. Results Four DNAzymes, named Dz1655, Dz1751, Dz1766 and Dz1789 were used for enzyme digestion. Results showed that only Dz1751 succeeded in cutting the full-length mRNA of ezrin at the expected sites, obtaining two fragments of 128 bp and 1 633 bp. After reaction, site AU1751 was found the least in the substrate RNA, which indicated that site AUI751 was the best target site for DNA- zymes. Conclusion Nucleic acid secondary structure combined with thermal kinetic parameters can predict and de- sign Ezrin mRNA target sites for DNAzymes.

关 键 词：EZRIN 靶点 DNAZYMES 二级结构 

分 类 号：R738[医药卫生—肿瘤]