Modifying the 5′-Cap for Click Reactions of Eukaryotic mRNA and To Tune Translation Efficiency in Living Cells
Modifying the 5′-Cap for Click Reactions of Eukaryotic mRNA and To Tune Translation Efficiency in Living Cells
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DOI：
10.1002/anie.201604107
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发表时间：
2016-08-26
影响因子：
16.6
通讯作者：
Rentmeister, Andrea
中科院分区：
文献类型：
--
作者：
Holstein, Josephin M.;Anhaeuser, Lea;Rentmeister, Andrea
5-帽是真核生物mRNA的标志，在RNA代谢中起着重要作用，从质量控制到输出和翻译。因此，修饰5-帽可以实现对潜在过程的调节和对几种生物功能的研究或调节。提出了一种简单的方法，用于基于高度混杂的甲基转移酶Ecm 1高效生产一系列N7修饰的帽。我们表明，这些，以及N-2-修饰的5-帽，可用于调整各自的mRNA的体外和细胞中的翻译。适当的修饰允许随后的生物正交化学，正如靶mRNA的细胞内活细胞标记所证明的那样。mRNA帽的有效和通用的N7操纵使得mRNA适合于它们的生物学功能和细胞内标记的调节，并且代表了对化学生物学工具箱的有价值的补充。
The 5-cap is a hallmark of eukaryotic mRNAs and plays fundamental roles in RNA metabolism, ranging from quality control to export and translation. Modifying the 5-cap may thus enable modulation of the underlying processes and investigation or tuning of several biological functions. A straightforward approach is presented for the efficient production of a range of N7-modified caps based on the highly promiscuous methyltransferase Ecm1. We show that these, as well as N-2-modified 5-caps, can be used to tune translation of the respective mRNAs both invitro and in cells. Appropriate modifications allow subsequent bioorthogonal chemistry, as demonstrated by intracellular live-cell labeling of a target mRNA. The efficient and versatile N7 manipulation of the mRNA cap makes mRNAs amenable to both modulation of their biological function and intracellular labeling, and represents a valuable addition to the chemical biology toolbox.