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。 Objective To construct the lentivirus vector containing Notch ligand Delta1 gene and establish human dental pulp stem cell line with stable interference of Delta1 gene. METHODS: Delta1 gene-specific siRNA target sequence was ligated with double-digested lentiviral vector pGCSIL-GFP, and the positive clones were identified by PCR. The recombinant lentivirus 293T was used to infect human dental pulp stem cells and divided into DPSCs / Delta1-RNAi group, DPSCs / vector group and DPSCs / wt group. The expression of Delta1 mRNA and protein were detected by RT-PCR and Western blot, respectively. Results The Delta1 specific siRNA lentiviral vector was successfully constructed and infected with dental pulp stem cells by PCR and DNA sequencing. The expression of Delta1 mRNA and protein in DPSCs / Delta1-RNAi group was significantly lower than that in DPSCs / vector group and DPSCs / wt group , There was no significant difference between DPSCs / vector group and DPSCs / wt group. Conclusion Transfection of human dental pulp stem cells with Delta1-specific siRNA lentiviral vectors successfully regulates Delta1 mRNA and protein expression.