| 小干扰RNA沉默胚胎生殖细胞Nanog基因的表达 |
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| 引用本文： | 王璐,何永林,李芳菲,彭彦,姜蓉,王亚平. 小干扰RNA沉默胚胎生殖细胞Nanog基因的表达[J]. 生殖与避孕, 2010, 30(1): 1-8 |
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| 作者姓名： | 王璐 何永林 李芳菲 彭彦 姜蓉 王亚平 |
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| 作者单位： | 1. 重庆医科大学组胚教研室,干细胞与组织工程研究室,重庆,400016
2. 重庆医科大学病原生物学教研室,重庆,400016 |
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| 基金项目： | 重庆医科大学科研基金(NSFYY200811、XBYB2007080)资助项目 |
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| 摘 要： | 目的:建立胚胎生殖细胞中沉默多能干细胞特异转录因子Nanog表达的方法。方法:通过检测碱性磷酸酶、SSEA-1和Oct4标志蛋白,鉴定体外培养的胚胎生殖细胞(EG细胞),通过脂质体介导作用,将Nanog特异性小干扰RNA(siRNA)转染EG细胞,应用半定量RT-PCR检测转染24h、36h、48h后EG细胞中Nanog mRNA的表达水平,并应用免疫荧光技术检测转染36h、48h后EG细胞中Nanog蛋白的表达水平。结果:培养细胞具有高度碱性磷酸酶活性,SSEA-1、Oct4阳性,为保持未分化状态的EG细胞。转染后Nanog mRNA和蛋白表达均受到抑制。结论:脂质体介导siRNA能有效沉默EG细胞中Nanog基因的表达,为进一步探讨Nanog对胚胎生殖细胞的调控机制奠定了基础。
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| 关 键 词： | 胚胎生殖细胞 小干扰RNA Nanog |
siRNA Induces Silence of Nanog Expression in Embryonic Germ Cells |
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| Lu WANG,Yong-lin HE,Fang-fei LI,Yan PENG,Rong JIANG,Ya-ping WANG. siRNA Induces Silence of Nanog Expression in Embryonic Germ Cells[J]. Reproduction and Contraception, 2010, 30(1): 1-8 |
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| Authors: | Lu WANG Yong-lin HE Fang-fei LI Yan PENG Rong JIANG Ya-ping WANG |
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| Affiliation: | Lu WANG1,Yong-lin HE2,Fang-fei LI1,Yan PENG1,Rong JIANG1,Ya-ping WANG1 (1 Department of Histology , Embryology,Lab of Stem Cell , Tissue Engineering,Chongqing,400016) (2. Department of Pathogens Biology,400016) |
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| Abstract: | Objective: To establish a method for silencing the expression of Nanog gene in embryonic germ cells. Methods: The embryonic germ cells (EG cells) cultured in vitro were identified by ALP staining via cytochemistry, SSEA-1 staining via immunocytochemistry and Oct4 staining via immunofluorescence. The EG cells were transfected by liposome-mediated small interfering RNA (siRNA), the level of Nanog mRNA was subsequently detected by semiquantitative RT-PCR at 24 h, 36 h, 48 h after transfection, respectively and... |
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| Keywords: | embryonic germ cell small interfering RNA Nanog |
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