| 孙懿1,2,3 卢光琇1,2,3 林戈1,2,3.利用逆转录病毒载体建立SMAD2 稳定干扰的人胚胎干细胞系[J].现代生物医学进展英文版,2012,12(15):2801-2803. |
| 利用逆转录病毒载体建立SMAD2 稳定干扰的人胚胎干细胞系 |
| Establishing SMAD2-Knocked down Human Embryonic Stem Cells byShRNA Retroviral Vector |
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| DOI： |
| 中文关键词: 人胚胎干细胞 RNA 干扰 SMAD2 |
| 英文关键词: Human embryonic stem cells RNAi SMAD2 |
| 基金项目:高等学校博士学科点专项科研基金新教师基金项目（200805331133）；国家"863" 计划项目（2006AA02A102）；国家
自然科学基金项目（81101510，30800659）；湖南省自然科学基金青年基金项目（09JJ4009）；中央高校基本科研业务费青年教师
助推基金项目（201012200219） |
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| 中文摘要: |
| 目的：建立SMAD2 稳定干扰的人胚胎干细胞系。方法：利用包装细胞获得重组的逆转录病毒，感染人类胚胎干细胞，为干扰
组ShSMAD2、载体组VECTOR 和野生型组WT；经荧光筛选获得阳性克隆；Realtime-PCR 检测SMAD2 mRNA 的表达。结果：带
GFP 荧光标记的SMAD2 特异性shRNA 逆转录病毒感染人类胚胎干细胞后，获得稳定干扰SMAD2 表达的人胚胎干细胞系。经
检测shSMAD2 组SMAD2 mRNA 表达较VECTOR 和WT 组明显降低，VECTOR 和WT 组之间无明显差异。结论：通过SMAD2
特异性shRNA 逆转录病毒载体构建了稳定干扰SMAD2 的人类胚胎干细胞。 |
| 英文摘要: |
| Objective: To establish stably SMAD2 shRNA interfered human embryonic stem cell line. Methods: SMAD2 shRNA
expressed pRetroSuper-GFP-SMAD2 vector was made virus in packaging cell line. Human embryonic stem cells were infected, which
were divided into interference group (ShSMAD2), VECTOR group and wild-type group (WT). Positive clones were selected by GFP
Fluorescence. SMAD2 knocked down efficiency were evaluated by Real-time PCR analysis. Results: After human embryonic stem cells
were infected with SMAD2-specific shRNA retrovirus labeled GFP fluorescence, stable interference of SMAD2 expression in human
embryonic stem cell lines had been obtained, in which SMAD2 mRNA expression was significantly lower compared with the VECTOR
and the WT group, while VECTOR group and WT group showed no significant difference. Conclusion: Retroviral vector-delivered
shRNA resulted in effective and stable down-regulation of SMAD2 expression in human embryonic stem cells. |
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