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Chemoenzymatic Preparation of Functional Click‐Labeled Messenger RNA
Author(s) -
Croce Stefano,
Serdjukow Sascha,
Carell Thomas,
Frischmuth Thomas
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201900718
Subject(s) - pseudouridine , messenger rna , click chemistry , translation (biology) , nucleotide , translational frameshift , chemistry , protein biosynthesis , rna , microbiology and biotechnology , biochemistry , biology , computational biology , combinatorial chemistry , uridine , gene
Synthetic mRNAs are promising candidates for a new class of transformative drugs that provide genetic information for patients’ cells to develop their own cure. One key advancement to develop so‐called druggable mRNAs was the preparation of chemically modified mRNAs, by replacing standard bases with modified bases, such as uridine with pseudouridine, which can ameliorate the immunogenic profile and translation efficiency of the mRNA. Thus the introduction of modified nucleobases was the foundation for the clinical use of such mRNAs. Herein we describe modular and simple methods to chemoenzymatically modify mRNA. Alkyne‐ and/or azide‐modified nucleotides are enzymatically incorporated into mRNA and subsequently conjugated to fluorescent dyes using click chemistry. This allows visualization of the labeled mRNA inside cells. mRNA coding for the enhanced green fluorescent protein (eGFP) was chosen as a model system and the successful expression of eGFP demonstrated that our modified mRNA is accepted by the translation machinery.