Figure 2.
Overview of in vitro transcription process and chemical modifications of IVT-mRNA used in this study
Main steps, and conditional post-transcriptional treatments, which were only applied to samples with cap modifications, i.e., methylation and dephosphorylation, are illustrated at the top panel. Simplified chemistry of IVT-mRNA synthesized and utilized at this study with various cap and/or nucleotide modifications are depicted at the bottom panel. Sequence of synthesis steps applied for cap modified mRNAs are indicated as colored bars corresponding to each synthesis step. Phosphatase treatment step is further indicated with/− or/+ next to sample name. Chemical formula only indicate the main variations among nucleotides as well as cap structures. See Figure S1 for precise chemical formula of cap and nucleotide modifications, and Table S1 for detailed information on chemistry and synthesis process of each sample. pDNA, plasmid DNA; T7 Pol., T7 RNA polymerase; NTs, nucleotides; ARCA, anti-reverse cap analog; Ψ, pseudouridine; me1Ψ, N1-Methylpseudouridine; 5moU, 5-methoxy-uridine; 5meC, 5-methyl-cytidine.