title: Synthetic mRNAs with superior translation and stability properties.
abstract: 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Lousisiana State University Health Sciences Center, Shreveport, LA, USA.
authors.forename: Ewa
authors.initials: E
authors.lastname: Grudzien-Nogalska
authors.forename: Joanna
authors.initials: J
authors.lastname: Kowalska
authors.forename: Wei
authors.initials: W
authors.lastname: Su
authors.forename: Andreas N
authors.initials: AN
authors.lastname: Kuhn
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Slepenkov
authors.forename: Edward
authors.initials: E
authors.lastname: Darzynkiewicz
authors.forename: Ugur
authors.initials: U
authors.lastname: Sahin
authors.forename: Jacek
authors.initials: J
authors.lastname: Jemielity
authors.forename: Robert E
authors.initials: RE
authors.lastname: Rhoads
mesh_terms: D000818:Animals; D006801:Humans; D014176:Protein Biosynthesis; D012314:RNA Cap Analogs / Q000737:chemistry*; D020871:RNA Stability; D012333:RNA, Messenger / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: D012314:RNA Cap Analogs; D012333:RNA, Messenger