靶向<i>hnRNP K</i>基因siRNA的筛选及其对精原细胞存活的影响

靶向hnRNP K基因siRNA的筛选及其对精原细胞存活的影响

摘要: 利用免疫细胞荧光、RNA干涉(RNA interfere,RNAi)、qRT-PCR、Western blot及细胞计数等方法,设计合成三对小分子干扰RNA (small interfering RNA,siRNA)(siRNA-hnRNP K-1,2,3),对小鼠精原细胞中内源性的不均一性核糖核蛋白K (Heterogeneous ribonucleoproteins K,hnRNP K)进行敲低并对其沉默效率进行筛选,研究其对小鼠精原细胞存活的影响.结果表明,siRNA-hnRNP K-3对小鼠精原细胞具有80%以上的沉默效率,转染后发现细胞数目明显减少,说明沉默hnRNP K基因的表达对小鼠精原细胞的存活有显著影响.

Abstract: Through the immune cell fluorescence, RNAi (RNA interfere), qRT-PCR, Western blot and cell counting methods, three pairs of siRNA (small interfering RNA) (siRNA-hnRNP K-1, 2, 3) were designed and synthesed to screen the silence efficiency that the endogenous hnRNP K of spermatogonium in mice were knockdown, and then studied the effect on spermatogonium survival. Results showed that the siRNA-hnRNP K-3 had silence efficiency more than 80%, and the cell number decreased significantly from further transfection to spermatogonium in mice. The results showed that the hnRNP K gene knockdown had a significant effect on the survival of spermatogonium in mice.