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Molecular composition and modification strategies for in vitro mRNA synthesis. (A) Several strategies have been applied to improve mRNA stability and translational efficiency, including cap optimization, nucleotide substitution and UTR-modifications. (B) In vitro synthesis of modified mRNA starts with plasmid cloning, followed by PCR amplification of the respective ORF. Lastly, cDNA is transcribed into mRNA, containing the optimized molecular composition.
