| 构建siRNA慢病毒载体调控人牙髓干细胞Delta1表达的实验研究 |
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| 引用本文： | 王雪飞,张纲,裘松波,何飞,谭颖微,汪蕾,吴小波. 构建siRNA慢病毒载体调控人牙髓干细胞Delta1表达的实验研究[J]. 第三军医大学学报, 2010, 32(1) |
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| 作者姓名： | 王雪飞 张纲 裘松波 何飞 谭颖微 汪蕾 吴小波 |
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| 作者单位： | 第三军医大学新桥医院口腔科,重庆,400037;第三军医大学新桥医院口腔科,重庆,400037;第三军医大学新桥医院口腔科,重庆,400037;第三军医大学新桥医院口腔科,重庆,400037;第三军医大学新桥医院口腔科,重庆,400037;第三军医大学新桥医院口腔科,重庆,400037;第三军医大学新桥医院口腔科,重庆,400037 |
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| 基金项目： | 重庆市自然科学基金(CSTC2007BB5016);;第三军医大学科研基金(2006D136)~~ |
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| 摘 要： | 目的构建Notch配体Delta1基因慢病毒干扰载体及建立Delta1基因稳定干扰的人牙髓干细胞系。方法将Delta1基因特异性siRNA靶序列与双酶切慢病毒载体pGCSIL-GFP连接,转化,挑取阳性克隆进行PCR鉴定;利用包装细胞293T获得重组的慢病毒,感染人牙髓干细胞,分为DPSCs/Delta1-RNAi组、DPSCs/vector组和DPSCs/wt组;RT-PCR和Western blot分别检测Delta1 mRNA及蛋白的表达。结果经PCR和DNA测序鉴定,成功构建Delta1特异性siRNA的慢病毒载体,并感染牙髓干细胞;经检测DPSCs/Delta1-RNAi组Delta1 mRNA及蛋白表达较DPSCs/vector组和DPSCs/wt组明显降低,DPSCs/vector组和DPSCs/wt组之间无明显差异。结论通过成功构建的Delta1特异性siRNA慢病毒载体对人牙髓干细胞的转染实现了对其Delta1 mRNA和蛋白的调控。
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| 关 键 词： | Dehtal Notch信号通路 RNA干扰 慢病毒 人牙髓干细胞 |
Regulation of Deltal expression in DPSCs by specific siRNA lentivirus vector |
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| Wang Xuefei,Zhang Gang,Qiu Songbo,He Fei,Tan Yinghui,Wang Lei,Wu Xiaobo. Regulation of Deltal expression in DPSCs by specific siRNA lentivirus vector[J]. Acta Academiae Medicinae Militaris Tertiae, 2010, 32(1) |
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| Authors: | Wang Xuefei Zhang Gang Qiu Songbo He Fei Tan Yinghui Wang Lei Wu Xiaobo |
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| Affiliation: | Department of Stomatology;Xinqiao Hospital;Third Military Medical University;Chongqing;400037;China |
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| Abstract: | Objective To construct a lentivirus vector for RNA interference(RNAi) of Delta1 gene and establish human dental pulp stem cells(DPSCs) with stably inhibited Delta1.Methods Delta1 specific siRNA was ligated with double digested lentivirus vector pGCSIL-GFP and the positive colonies were obtained by PCR,DPSCs were interfered and divided into DPSCs/Delta1-RNAi group,DPSCs/vector group and DPSCs/wt group,and the expression of Delta1 was verified by RT-PCR and Western blot analysis.Results The ligation of Delta1... |
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| Keywords: | Delta1 notch signal siRNA lentivirus human dental pulp stem cells |
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