Westerich Kim J, Chandrasekaran Karthik S, Gross-Thebing Theresa, Kueck Nadine, Raz Erez, Rentmeister Andrea
Institute of Cell Biology Center for Molecular Biology of Inflammation , University of Münster , D-48149 Münster , Germany . Email:
Institut für Biochemie , Westfälische Wilhelms-Universität Münster , Wilhelm-Klemm-Str. 2 , 48149 Münster , Germany . Email:
Chem Sci. 2020 Feb 24;11(11):3089-3095. doi: 10.1039/c9sc05981d. eCollection 2020 Mar 21.
Live imaging of mRNA in cells and organisms is important for understanding the dynamic aspects underlying its function. Ideally, labeling of mRNA should not alter its structure or function, nor affect the biological system. However, most methods applied make use of genetically encoded tags and reporters that significantly enhance the size of the mRNA of interest. Alternately, we utilize the 3' poly(A) tail as a non-coding repetitive hallmark to covalently label mRNAs bioorthogonal chemistry with different fluorophores from a wide range of spectra without significantly changing the size. We demonstrate that the labeled mRNAs can be visualized in cells and zebrafish embryos, and that they are efficiently translated. Importantly, the labeled mRNAs acquired the proper subcellular localization in developing zebrafish embryos and their dynamics could be tracked .
对细胞和生物体中的信使核糖核酸（mRNA）进行实时成像，对于理解其功能背后的动态过程至关重要。理想情况下，mRNA的标记不应改变其结构或功能，也不应影响生物系统。然而，大多数应用的方法利用了基因编码标签和报告基因，这会显著增加目标mRNA的大小。另外，我们利用3' 聚腺苷酸（poly(A)）尾作为非编码重复特征，通过生物正交化学方法用来自广泛光谱范围的不同荧光团共价标记mRNA，而不会显著改变其大小。我们证明，标记的mRNA可以在细胞和斑马鱼胚胎中可视化，并且它们能够高效翻译。重要的是，标记的mRNA在发育中的斑马鱼胚胎中获得了正确的亚细胞定位，并且可以追踪它们的动态变化。
