mRNA Synthesis: Overcoming Challenges of In Vitro Transcription (IVT) DNA template preparation with Long Poly(A) Sequences

mRNA Synthesis: Overcoming Challenges of In Vitro Transcription (IVT) DNA template preparation with Long Poly(A) Sequences

Synthetic mRNA templates are widely used for in vitro transcription (IVT) and have become essential tools for mRNA research, vaccine development, and gene therapy. A crucial component of every IVT plasmid is the poly(A) tail, which plays a central role in mRNA stability and efficient protein expression. While longer poly(A) tails, typically 100 to 150 bases, enhance mRNA stability, they are also prone to truncation during plasmid propagation in E. coli. These shortened or heterogeneous poly(A) segments can compromise mRNA performance, reduce stability, and generate inconsistent expression results.

To address these challenges, GENEWIZ from Azenta Life Sciences has developed an optimized plasmid preparation protocol that maintains the integrity of long poly(A) sequences of at least 140 bases. Using a proprietary E. coli strain and refined culturing conditions, this workflow stabilizes extended poly(A) regions, increases plasmid yield, and produces high-fidelity DNA templates ideal for IVT and downstream mRNA synthesis.

What you'll learn:

• How synthetic mRNA plasmids are designed and prepared for in vitro transcription
• Why long poly(A) tails are essential for mRNA stability and protein expression
• Key challenges associated with producing plasmids containing poly(A) sequences >100 bases
• Performance comparison between standard plasmid preparation methods and GENEWIZ’s optimized protocol
• How a proprietary E. coli strain improves poly(A) integrity, reduces truncation, and boosts mega-prep DNA yield