中国科学技术大学学报 ›› 2016, Vol. 46 ›› Issue (10): 797-802.DOI: 10.3969/j.issn.0253-2778.2016.10.001

• 原创论文 •    下一篇

正交点击反应对应用于标记细胞器

罗宇峰   

  1. 中国科学技术大学化学系,安徽合肥 230026
  • 收稿日期:2016-04-20 修回日期:2016-07-05 出版日期:2016-10-31 发布日期:2016-10-31
  • 通讯作者: 梁高林
  • 作者简介:罗宇峰,男,1992年生,硕士. 研究方向:分子影像学. E-mail: swlyf@mail.ustc.edu.cn
  • 基金资助:
    国家自然科学基金联合基金(U1532144)资助.

Labelling cell organelles with dual orthogonal click reaction pairs

LUO Yufeng   

  1. Department of Chemistry, University of Science and Technology of China, Hefei 230026, China
  • Received:2016-04-20 Revised:2016-07-05 Online:2016-10-31 Published:2016-10-31

摘要: 单一点击反应已广泛应用于细胞成像,但是利用多个正交反应对同一生物体系进行多重标记仍然是一个挑战.本研究将两种不同机理的点击反应——一种是tetrazine-alkene [4+2] Diels-Alder cycloaddition(四嗪-烯烃[4+2] 狄尔斯-阿尔德环加成反应),另外一种是CBT-Cys condensation reaction(2-氰基苯并噻唑-半胱氨酸缩合反应)——结合起来,验证其正交性,并将其运用于常见的HepG2细胞的细胞器成像中,实现了同时显示细胞线粒体和细胞膜的状态.实验结果说明在同一细胞中同时运用两个(或者多个)点击反应标记多个细胞器是可行的.

关键词: 点击反应, 正交性, 细胞成像

Abstract: While single click reactions for cell imaging are widely applied, employing multiple orthogonal click reactions tracking multiple complex intracellular systems within one cell remains challenging. Our research combined two click reactions with different reaction mechanisms: tetrazine- alkene [4+2] inverse electron demand Diels-Alder cycloaddition, and CBT-Cys condensation reaction. Orthogonality between these two reactions was studied and the two reactions were applied for simultaneous labeling mitochondria and outer cell membrane of HepG2 cells. The results indicate that it is possible to use two (or n) orthogonal click reactions to track multiple organelles within one cell.

Key words: click reaction, orthogonality, cell imaging