6533b7dcfe1ef96bd127290b

RESEARCH PRODUCT

Synthetic mRNAs with Superior Translation and Stability Properties

Robert E. RhoadsEdward DarzynkiewiczSergey V. SlepenkovAndreas KuhnJoanna KowalskaWei SuJacek JemielityEwa Grudzien-nogalskaUgur Sahin

subject

RNA StabilityMessenger RNAchemistry.chemical_compoundRNA Cap AnalogsTranslational efficiencyChemistryRNA polymeraseProtein biosynthesisRNAContext (language use)Cell biology

description

The translational efficiency and stability of synthetic mRNA in both cultured cells and whole animals can be improved by incorporation of modified cap structures at the 5'-end. mRNAs are synthesized in vitro by a phage RNA polymerase transcribing a plasmid containing the mRNA sequence in the presence of all four NTPs plus a cap dinucleotide. Modifications in the cap dinucleotide at the 2'- or 3'-positions of m(7)Guo, or modifications in the polyphosphate chain, can improve both translational efficiency and stability of the mRNA, thereby increasing the amount and duration of protein expression. In the context of RNA-based immunotherapy, the latter is especially important for antigen production and presentation by dendritic cells. Protocols are presented for synthesis of modified mRNAs, their introduction into cells and whole animals, and measurement of their translational efficiency and stability.

yearjournalcountryeditionlanguage
2012-11-08
https://doi.org/10.1007/978-1-62703-260-5_4