PolyA tail segmentation improves the stability of the template DNA and increases the translatability of in vitro transcribed mRNA.

ABSTRACT: PolyA tail regulates messenger RNA (mRNA) localization, stability, and translation. PolyA length affects the durability and translational activity of both endogenously and exogenously delivered mRNAs. However, long polyA stretches can undergo recombination during amplification in bacterial plasmids, impairing the production of in vitro transcribed mRNA with long polyAs. PolyA tail segmentation with heteronucleotide spacers has recently emerged as a solution. Here, we developed segmented polyA patterns that stabilize the sequence during DNA amplification and enhance mRNA translation. We designed 15 novel genetically modified polyA variants, differing in the length, placement, and frequency of spacers, and the overall length (from ~120 to 200 nucleotides). We evaluated their stability in DNA

SUBMITTER: Spiewla T 

PROVIDER: S-EPMC12805897 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature