基于CIRP基因有效干扰靶点的筛选

Screening based on the effective interference target of the cold- inducible RNA- binding protein( CIRP) gene

为了研究冷诱导RNA结合蛋白(CIRP)更深入的功能及意义,试验通过Lipo FiterTM将黑龙江八一农垦大学动物生理生化实验室之前已构建好的5组RNA干扰重组载体及空载体分别瞬时转染到大鼠肝细胞中,提取细胞RNA经反转录后进行荧光定量PCR试验,获得各组干扰载体对CIRP基因干扰后CIRP基因mRNA的相对表达量,确定效果最佳的干扰靶点。结果表明:在5组干扰试验组CIRP基因mRNA的表达量中,264组与空白对照组的表达量差异显著(P<0.05),并且干扰效率达到70%以上;阴性对照组(空载体组)与空白对照组的表达量基本相同,差异不显著(P>0.05)。说明试验成功筛选出有效干扰靶点。

To study the in -depth function and significance of the cold -inducible RNA -binding protein （CIRP）, the RNA interference vectom in the five groups and empty vectors constructed by the laboratory of animal physiology and biochemistry of Heilongjiang Bayi agricultural university, were transiently transfected into the rat hepatocytes by LipoFiterTM, respectively. The RNA extracted from the cells, was used for fluorescence quantitative PCR experiments after reverse transcription. The relative expression levels of CIRP mRNA after interference with the interference vectors were obtained to determine the best interference targets. The results showed that, among the expression levels of CIRP mRNA in the five interference experimental groups, there was a significant difference （ P 〈 0.05 ） in the expression levels between the 264 group and the blank control group, and the efficiency of interference reached more than 70% ; there was no significant difference （ P 〉 0.05 ） in the expression levels between the negative control group （ empty vector） and the blank control group due to the similar expression levels. The results indicate that the effective interference targets are successfully screened.