高效能沉默BMP-7基因siRNA序列的实验筛选

发布时间：2018-10-11 20:07

【摘要】：目的骨形态发生蛋白(bone morphogenetic protein-7,BMP-7)是一种促进骨和软骨再生的关键性生长因子,旨在筛选高效能BMP-7si RNA序列,为后续探讨BMP-7的生物学作用机制的基础和临床应用研究提供实验研究技术手段。方法本实验针对BMP-7基因,构建出相应4条si RNA序列,分别将其转染至大鼠脐静脉内皮细胞,转染72 h后用Wb和RT-PCR方法检测BMP-7蛋白和BMP-7 m RNA表达水平,筛选出最高效的BMP-7si RNA序列。结果人工合成的4条BMP-7si RNA序列都能有效抑制大鼠脐静脉内皮细胞的BMP-7表达,其中BMP-7-3si RNA序列抑制效果最为明显,抑制率大于90%,被确认为体外抑制BMP-7表达最为有效的干扰序列。结论 BMP-7-3 si RNA序列能够在体外高效能抑制BMP-7表达,故可作为一种实验研究技术手段,用于探讨BMP-7的生物学作用机制的基础和临床应用研究。
[Abstract]:Objective Bone morphogenetic protein (bone morphogenetic protein-7,BMP-7) is a key growth factor to promote bone and cartilage regeneration. To provide experimental techniques for the further study of the biological mechanism and clinical application of BMP-7. Methods four si RNA sequences of BMP-7 gene were constructed and transfected into rat umbilical vein endothelial cells. After 72 h of transfection, the expression levels of BMP-7 protein and BMP-7 m RNA were detected by Wb and RT-PCR methods, and the most efficient BMP-7si RNA sequences were screened out. Results all of the four synthetic BMP-7si RNA sequences could effectively inhibit the expression of BMP-7 in rat umbilical vein endothelial cells. The inhibitory rate of BMP-7-3si RNA sequence was more than 90, which was confirmed as the most effective interference sequence to inhibit the expression of BMP-7 in vitro. Conclusion BMP-7-3 si RNA sequence can effectively inhibit the expression of BMP-7 in vitro, so it can be used as an experimental technique to study the biological mechanism and clinical application of BMP-7.
【作者单位】：昆明医科大学;昆明医科大学第一附属医院;
【基金】：国家自然科学基金(81160239,81460296) 云南省科技厅-昆明医科大学联合专项重点项目(2012FB161)~~
【分类号】：R3416

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