组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2012, Vol. 8 ›› Issue (2): 69-72.doi: 10.3969/j.issn.1673-0364.2012.02.003

• 论著 • 上一篇    下一篇

角质形成细胞PAR-2基因SIRNA的筛选

袁磊，金蓉，张艳，张余光   

  1. 上海交通大学医学院附属第九人民医院整复外科
  • 发布日期:2020-07-23

Screening of siRNA Targeted PAR-2 in Keratinocyte

YUAN Lei,JIN Rong,ZHANG Yan,ZHANG Yuguang   

  1. Department of Plastic and Reconstructive Surgery,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200011,China.
  • Published:2020-07-23
  • Contact: 上海市教育委员会科研项目　　（06BZ021）

摘要： 目的设计并筛选能高效抑制人角质形成细胞膜上蛋白酶激活受体-2(PAR-2)基因表达的小干扰RNA(siRNA)序列。方法根据人PAR-2 mRNA的序列,设计合成3对针对靶基因的不同siRNA。应用脂质体Lipofectamin2000转染试剂,将3对siRNA转染人角质细胞,半定量RT-PCR技术检测PAR-2mRNA表达水平、Western blot检测PAR-2蛋白表达,以评价干扰效果。结果设计合成的3对PAR-2 siRNA中有2对siRNA可有效抑制PAR-2基因表达,以第2对siRNA效果为佳。结论成功设计合成特异且高效阻断PAR-2表达的siRNA,为进一步应用RNAi技术沉默PAR-2基因奠定实验基础。

关键词: 角质形成细胞, 蛋白酶激活受体-2, RNA干扰

Abstract: Objective To design and screen small interfering RNA(siRNA) targeted protease-activated receptor-2(PAR-2) mRNA in Keratinocyte that can specifically and effectively suppress PAR-2 expression.Methods Three pairs of PAR-2 specific double stranded siRNAs were designed according to the sequence of PAR-2 mRNA.These siRNA were transfected into human keratinocyte by Lipofectamine 2000.The transcription level of PAR-2 was analyzed by semi-quantitative RT-PCR and PAR-2 protein expression was tested by western blot in order to evaluate the effect of RNA interference.Results Two of the three PAR-2 siRNA effectively inhibited the expression of PAR-2 and the second one is more effective.Conclusion PAR-2 siRNAs can be successfully designed and synthesized,which can specifically and effectively suppresses the expression of PAR-2 gene.The results in this study lay the foundation for further study on PAR-2 gene silencing by RNA interfere(RNAi).

Key words: Keratinocyte, Protease-activated receptor-2, RNA interfere

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