[1]王雪飞,张纲,裘松波,等.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(01):30-32.
 Wang Xuefei,Zhang Gang,Qiu Songbo,et al.Regulation of Delta1 expression in DPSCs by specific siRNA lentivirus vector[J].J Third Mil Med Univ,2010,32(01):30-32.
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构建siRNA慢病毒载体调控人牙髓干细胞Delta1表达的实验研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
32卷
期数:
2010年第01期
页码:
30-32
栏目:
论著
出版日期:
2010-01-15

文章信息/Info

Title:
Regulation of Delta1 expression in DPSCs by specific siRNA lentivirus vector
作者:
王雪飞张纲裘松波何飞谭颖徽汪蕾吴小波
第三军医大学新桥医院口腔科
Author(s):
Wang Xuefei Zhang Gang Qiu Songbo He Fei Tan Yinghui Wang Lei Wu Xiaobo
Department of Stomatology, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China
关键词:
Delta1Notch信号通路RNA干扰慢病毒人牙髓干细胞
Keywords:
Delta1 notch signal siRNA lentivirus human dental pulp stem cells
分类号:
R322.41;R329.24;R394.1
文献标志码:
A
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。
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 specific siRNA to the double digested lentivirus vector pGCSIL-GFP was successful. Delta1 protein and Delta1 mRNA expression of DPSCs/Delta1-RNAi group was significantly higher than the DPSCs/vector group and DPSCs/wt group, but there was no different between the DPSCs/vector group and DPSCs/wt group. Conclusion  The construction of Delta1 specific siRNA lentivirus vectors is successful and it regulates the expression of Delta1 in DPSCs

参考文献/References:

王雪飞, 张纲, 裘松波, 等.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):30-32.

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更新日期/Last Update: 2010-01-08