| 大鼠海马神经干细胞Brn-4基因的特异性siRNA研究 |
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| 引用本文： | 朱蕙霞,施金洪,金国华,田美玲,秦建兵.大鼠海马神经干细胞Brn-4基因的特异性siRNA研究[J].苏州大学学报(自然科学版),2009,29(6):1021-1025. |
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| 作者姓名： | 朱蕙霞 施金洪 金国华 田美玲 秦建兵 |
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| 作者单位： | 朱蕙霞,ZHU Hui-xia(南通大学医学院,生物化学教研室,江苏南通,226001);施金洪,金国华,田美玲,秦建兵,SHI Jin-hong,JIN Guo-hua,TIAN Mei-ling,QIN Jian-bing(南通大学医学院,人体解剖学教研室,神经生物学研究所,江苏省神经再生重点实验室,江苏南通,226001) |
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| 基金项目： | 国家自然科学基金，江苏省自然科学基金 |
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| 摘 要： | 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。
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| 关 键 词： | 海马 神经干细胞 siRNA Brn-4 大鼠 |
Study on Specific siRNA for Brn-4 in Neural Stem Cells from Rat Hippocampus |
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| ZHU Hui-xia,SHI Jin-hong,JIN Guo-hua,TIAN Mei-ling,QIN Jian-bing.Study on Specific siRNA for Brn-4 in Neural Stem Cells from Rat Hippocampus[J].Suzhou University Journal of Medical Science,2009,29(6):1021-1025. |
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| Authors: | ZHU Hui-xia SHI Jin-hong JIN Guo-hua TIAN Mei-ling QIN Jian-bing |
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| Institution: | ZHU Hui-xia, SHI Jin-hong, JIN Guo-hua, TIAN Mei-ling, QIN Jian-bing (1. Dept of Biochemistry,2. Dept of Human Anatomy, Institute of Neurobiology,Jiangsu Key Laboratory of Neuroregeneration, Medical School, Nantong University, Jiangsu Nantong 226001, China) |
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| Abstract: | Objective To screen high effective and specific siRNA for silencing Brn-4 gene in rat hippocampal neural stem cells（NSCs）. Methods Four pairs of specific siRNAs for suppressing Brn-4 expression were designed and transfected into rat hippocampal NSCs in vitro by means of lipidosome. Brn-4 expression in transfection cells was detected by the methods of RT-PCR and immunofluoreseence assay, which were used for screening out the most effective siRNA for silencing Brn-4 gene and optimizing transfection condition. Results Brn-4 expression in rat hippocampal NSCs at different levels was suppressed. By four pairs of specific siRNAs I^# siRNA had the highest suppression efficiency,whose optimum transfection concentration was 0.5 μmol/L and optimum transfection time was 48 hours. Conclusion Applying I^# siRNA can silence Brn-4 expression in rat hippocampal NSCs with high efficiency. |
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| Keywords: | siRNA Brn-4 |
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