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为解决siRNA药物应用过程中存在的siRNA筛选和体内转运的问题,以乙肝病毒基因HBx为研究对象,通过构建表达目的基因和荧光素酶融合蛋白的双荧光素酶质粒,体外筛选得到有效抑制HBx基因的表达siRNA序列的质粒,并将该siRNA表达质粒装载于PEG化修饰的新型阳离子脂质体中,在细胞和动物水平开展该阳离子脂质体的作用效果研究。结果表明,上述装载siRNA质粒的阳离子脂质体能够在细胞和动物水平有效地抑制乙肝病毒HBx基因的表达,解决了siRNA应用中序列筛选和体内转运的问题。
In order to solve the problem of siRNA screening and translocation in the process of siRNA drug application, HBx gene of hepatitis B virus was used as the research object. The double luciferase plasmid expressing the target gene and the luciferase fusion protein was screened in vitro to effectively inhibit HBx Gene expression of the siRNA sequence of the plasmid, and the siRNA expression plasmid was loaded in pegylated modified novel cationic liposomes, the cellular and animal level to carry out the role of the cationic liposome effect study. The results showed that the above cationic liposomes loaded siRNA plasmid can effectively inhibit the expression of hepatitis B virus HBx gene at the level of cells and animals and solved the problem of sequence selection and in vivo transport in siRNA applications.