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目的筛选出高效针对大鼠海马神经干细胞Brn-4基因沉默的特异性siRNA。方法设计4对抑制Brn-4基因表达的特异性siRNA序列,通过脂质体转染入体外培养的大鼠海马神经干细胞中,采用RT-PCR和免疫荧光检测转染细胞Brn-4基因的表达,筛选出沉默Brn-4基因最为有效的siRNA序列,并优化其转染条件。结果4对siRNA不同程度地阻断了海马神经干细胞Brn-4基因的表达,阻断效率最高的1~# siRNA最适转染剂浓度为0.5μmol/L,最佳转染时长为48 h。结论应用1~# siRNA可以高效率地沉默大鼠海马神经干细胞Brn-4基因的表达。
Objective To screen highly specific siRNA targeting Brn-4 gene silencing in rat hippocampal neural stem cells. Methods Four siRNAs targeting Brn-4 gene were designed and transfected into rat hippocampus neural stem cells by lipofectamine. The expression of Brn-4 gene was detected by RT-PCR and immunofluorescence. , The most effective siRNA sequence of Silencing Brn-4 gene was screened and its transfection conditions were optimized. Results Four siRNAs blocked the expression of Brn-4 gene in hippocampal neural stem cells to different extents. The highest transfection efficiency of 1 ~ # siRNA was 0.5 μmol / L and the optimal transfection time was 48 h. Conclusion The expression of Brn-4 gene in rat hippocampus neural stem cells can be efficiently silenced by 1 ~ # siRNA.