生物学杂志

生物学杂志 ›› 2021, Vol. 38 ›› Issue (2): 26-.doi: 10.3969/j.issn.2095-1736.2021.02.026

• 研究报告 • 上一篇    下一篇

石墨烯量子点提高DNAzyme在细胞水平上的稳定性

  

  1. 1.华东理工大学生物反应器工程国家重点实验室；2.华东理工大学药学院，上海200237
  • 出版日期:2021-04-18 发布日期:2021-04-25
  • 通讯作者: 张井岩，博士，教授，研究方向为生物化学和生物无机化学， E⁃mail：zhangjy2015@gmail.com； 郑静，博士，副教授，研究方向为分子生物学、肿瘤生物学等， E⁃mail：zhengjing@ecust.edu.cn
  • 作者简介:黄奇萍，硕士，研究方向为DNAzyme活性研究， E⁃mail：1925260880@qq.com
  • 基金资助:
    国家自然科学基金（21671065）；中央高校基本科研业务费专项资金

Graphene quantum dots improve the stability of DNAzyme in cells

  1. 1.State⁃Key Laboratory of Bioreactor Engineering；2.School of Pharmacy，East China University of Science and Technology，Shanghai 200237，China
  • Online:2021-04-18 Published:2021-04-25

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摘要： 为了提高脱氧核糖核酸酶（又称DNAzyme）的稳定性，以石墨烯量子点（GQDs）为载体，负载两个靶向TNF⁃α基因的DNAzyme，考察该体系在分子和细胞水平抑制TNF⁃α蛋白表达的能力。体外分子水平的研究发现，DNAzyme与GQDs结合后，不仅保持了原有的DNAzyme活性，而且稳定性得到了提高。结果初步表明GQDs可以作为DNAzyme的载体。细胞水平的进一步研究发现，该体系与Hela细胞共培育36 h后，TNF⁃α蛋白水平表达量降低了50%，而单独DNAzyme作用的细胞中TNF⁃α蛋白表达量则基本不变，这表明GQDs可以将该DNAzyme载入细胞内，并提高了DNAzyme在细胞内的活性。结合分子水平的实验结果，初步推测DNAzyme活性的提高是因为GQDs载体的作用而使它在细胞水平的稳定性得到改善。

关键词: 脱氧核糖核酸酶, 石墨烯量子点, 稳定性, TNF?α

Abstract: In this work，we employed Graphene quantum dots（GQDs）as a carrier loading two RNA⁃cleaving Deoxyribonuclease（DNAzymes）to explore their activities at molecular and cellular levels. The two DNAzymes were designed and synthesized based on the mRNA of TNF⁃α（TNF⁃DNAzyme1 and TNF⁃DNAzyme2）. We demonstrated that the TNF⁃DNAzymes／GQDs conjugates were stable in solution，and the catalytic activity of the DNAzymes remained in the presence of GQDs. The expression level of TNF⁃α in Hela cells was significantly decreased after the incubation with TNF⁃DNAzymes and GQDs. In contrast，the expression level of TNF⁃α in Hela cells was barely changed after the incubation with TNF⁃DNAzymes alone，which indicated that GQDs could be used as a carrier to deliver TNF⁃DNAzymes into cells，thus the activity of TNF⁃DNAzymes in cellular level could be greatly improved. Based on the control experiments，we inferred that the improved activity of DNAzymes could be due to their improved stability with GQDs. However，we could not rule out which caused by the higher accumulation of DNAzymes in the cells or other mechanisms

Key words: DNAzyme, graphene quantum dots, stability, TNF?α

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