Modifying the 5′‐Cap for Click Reactions of Eukaryotic mRNA and To Tune Translation Efficiency in Living Cells |
| |
| Authors: | Josephin M Holstein Lea Anhäuser Prof?Dr Andrea Rentmeister |
| |
| Affiliation: | 1. University of Muenster, Department of Chemistry, Institute of Biochemistry, Muenster, Germany;2. Cells-in-Motion Cluster of Excellence (EXC 1003—CiM), University of Muenster, Germany |
| |
| Abstract: | 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 N2‐modified 5′‐caps, can be used to tune translation of the respective mRNAs both in vitro 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. |
| |
| Keywords: | bioorthogonal chemistry Ecm1 mRNA RNA labeling translation |
|
|
