Figure 1.

Plasmid template for generation of synthetic modified RNAs. Structural diagram shows “ready-to-use” plasmid containing 5′ and 3′ UTR sequences used to generate smRNAs of interest. The ORFs for PDX1, NGN3, and MAFA were cloned and subsequently excised outwardly to the UTR regions using Acc65I and SphI enzymes. A polyA tail was added to the template with a polymerase chain reaction, and the amplicons were then used for in vitro transcription. Abbreviations: ORF, open reading frame; pUC, origin of replication; smRNA, synthetic modified mRNA; UTR, untranslated region.