| 小鼠STAT3基因干扰质粒的构建及其效果鉴定 |
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| 引用本文： | 星懿展,李泽桂,李红丽,李铁石,丁震宇. 小鼠STAT3基因干扰质粒的构建及其效果鉴定[J]. 第三军医大学学报, 2006, 28(10): 1022-1025 |
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| 作者姓名： | 星懿展 李泽桂 李红丽 李铁石 丁震宇 |
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| 作者单位： | 第三军医大学基础医学部组织胚胎学教研室,重庆市神经科学研究所,重庆,400038;清华大学生命科学与医学研究院基因组研究所,北京,100084 |
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| 基金项目： | 中国科学院资助项目，重庆市人口计生委科研项目 |
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| 摘 要： | 目的构建小鼠STAT3基因的特异性RNA干扰质粒,并鉴定其在细胞水平的抑制效果.方法采用DNA载体介导的RNA干扰技术构建小鼠STAT3基因的特异性RNAi质粒(pBS/U6/STAT3-siRNA),并通过RT-PCR、免疫印迹、免疫细胞化学染色等技术对pBS/U6/STAT3-siRNA的抑制效率进行鉴定.结果经双酶切鉴定筛选出4个阳性干扰质粒,分别转入293T细胞后均可明显降低STAT3的表达,尤以靶向( 2 334～ 2 351)位点的质粒干扰效率最高,达70%.结论小鼠STAT3基因的特异性干扰质粒(pBS/U6/STAT3-siRNA)能从蛋白及mRNA水平上特异和高效地抑制STAT3基因的表达.这种DNA载体介导的双链RNA干扰质粒为进一步探究STAT3与胚胎发育的相关功能提供一种有效的研究工具.
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| 关 键 词： | pBS/U6/STAT3-siRNA STAT3 RNAi 小鼠 |
| 文章编号： | 1000-5404（2006）10-1022-04 |
| 收稿时间： | 2005-12-14 |
| 修稿时间： | 2006-03-28 |
Construction and suppressive effect of siRNA targeting mouse STAT3 |
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| XING Yi-zhan,LI Ze-gui,LI Hong-li,LI Tie-shi,DING Zhen-yu. Construction and suppressive effect of siRNA targeting mouse STAT3[J]. Acta Academiae Medicinae Militaris Tertiae, 2006, 28(10): 1022-1025 |
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| Authors: | XING Yi-zhan LI Ze-gui LI Hong-li LI Tie-shi DING Zhen-yu |
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| Affiliation: | 1. Department of Histology and Embryology, College of Medicine, Third Military Medical University, Chongqing 400038; 2.Institute of Biomedicine, Tsinghua University, Beijing 100084, China |
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| Abstract: | Objective To construct the specific interference plasmid targeting mouse STAT3 gene, and identify its suppressive effect. Methods The specific siRNAs targeting STAT3 gene was constructed. The semi-quantificational RT-PCR,Western blotting, immunocytochemical staining, cell proliferation and luciferase assay were used to confirm the validity of the recombinant siRNA plasmid. Results The siRNA plasmid was proved specifically to block the activation of over-expressed STAT3 in a dose-dependent manner in 293T cell, not only at protein level but also at mRNA level. Conclusion RNAi could specifically and efficiently inhibit STAT3 gene expression, and it will become an effective tool for searching the function of STAT3 relating to embryogenesis. |
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| Keywords: | pBS/U6/STAT3-siRNA STAT3 RNAi |
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