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[目的]构建肿瘤坏死因子-α诱导蛋白-1 (TNFAIP1)基因短发卡 RNA (shRNA)慢病毒载体.[方法]根据人TNFAIP1 基因序列设计合成4条 shRNA,构建 shRNA表达质粒,将重组质粒转染至 293T细胞,应用 RT-PCR 检测各组 TNFAIP1 基因 mRNA 的表达水平,筛选出最有效的干扰靶序列,构建HSH018143-LVRH1P-RNAi慢病毒载体,并进行测序鉴定.将确定了的 TNFAIP1-shRNA-0S523409 转染至293T细胞,72 h后,观察绿色荧光表达情况,并测定病毒滴度.[结果]TNFAIP1-shRNA慢病毒表达载体高效转导入 293T细胞并稳定表达,测定其病毒滴度为5×108TU/mL.[结论]本研究成功构建了TNFAIP1基因 shRNA慢病毒表达载体,为研究TNFAIP1 在糖尿病视网膜病变中的作用奠定了实验基础.“,”[Objective]To construct tumor necrosis factor alpha-induced protein 1 (TNFAIP1 )-shRNA lentiviral vector.[Methods]Four shRNA sequences were designed and synthesized according to the sequence of human TNFAIP1 gene.These recombinant plasmids were transfected into 293T cells.RT-PCR assay was ap-plied to detect the mRNA expression level of TNFAIP1 in each group and thus to choose the most effective TNFAIP1-shRNA.The HSH018143-LVRH1P-RNAi was constructed and confirmed by DNA sequencing. The selected TNFAIP1-shRNA-0S523409 was transfected into 293T cells.After 72 hours,the green fluores-cence expression was observed and the lentiviral titer was tested.[Results]The TNFAIP1-shRNA lentiviral vector had been transfected into 293T cells efficiently and stably,the titer of lentivirus was 5×108TU/mL.[Conclusion]The TNFAIP1-shRNA lentiviral expression vector has been constructed successfully,which lays a foundation for exploring for the role of TNFAIP1 in diabetic retinopathy.