抑制A549细胞COX-2表达的D-siRNAs的合成

目的:利用长片段双链RNA(dsRNAs)在体外经Dicer酶消化获得小RNA(D-siRNAs)的方法合成抑制A549细胞COX-2表达的siRNAs,以期获得治疗气道炎症和肿瘤的一种新思路.方法:用RiboMAX Large Scale RNA Production Systems-SP6 and T7试剂盒体外将COX-2 DNA转录为RNA.长片段COX-2 RNA经Dicer酶消化为小RNA,纯化并获取小RNA(D-siRNAs).结果:在体外,长片段双链A549细胞COX-2的RNA经Dicer酶消化后可成功获得抑制A549细胞COX-2表达的小RNA.结论:在体外,长片段dsRNAs经Dicer酶消化可成功获得21～23个碱基的小RNA.

Dicer efficiently converts large dsRNAs intosiRNAs suitable for COX-2 gene

OBJECTIVE: To report the use of Dicer to cleave double-stranded RNA (dsRNAs) into small interference RNAs (D-siRNAs) that can target multiple sites within an mRNA, and to acquire an new method to cure inflammation of the airway and tumor. METHODS: Using RiboMAX Large Scale RNA Production Systems-SP6 and T7 kit were used to transcribe A549 cell COX-2 DNA into RNA (dsRNAs). We mixed dsRNAs with Dicer in the reaction buffer. We recovered siRNAs using RNA Purification Column. RESULTS: Dicer efficiently converted double-stranded RNA of COX-2 into small interference RNAs of 21 approximately 23 bp. CONCLUSION: Dicer efficiently converts double-stranded RNA (dsRNA) into small interference RNAs (D-siRNAs of 21 approximately 23 bp).