RNA Molecule

Reducing Double-Stranded RNA Formation in In Vitro Transcription using a Novel RNA Polymerase

Double-stranded RNA (dsRNA) is a significant contaminant in mRNA. During in vitro transcription (IVT), the RNA polymerase transcribes template DNA to produce the desired RNA for downstream applications, such as a vaccine or other therapeutics.

During IVT, RNA polymerases naturally generate dsRNA by products through a number of mechanisms including cryptic promoters or loopback RNAs.

These dsRNA contaminants can then trigger innate immune activation and diminish protein expression, thereby jeopardizing the safety, tolerability, and efficacy of the potential therapeutic product. 

TriLink BioTechnologies has introduced a novel RNA polymerase, CleanScribe™ RNA Polymerase, that reduces dsRNA formation by up to 85% with simple integration into IVT for seamless mRNA synthesis.

CleanScribe™ RNA Polymerase is a novel DNA-dependent RNA polymerase that catalyzes the in vitro transcription (IVT) of a recombinant gene regulated by the T7 promoter.

10% Off CleanScribe RNA Polymerase Orders!

Enjoy reduced pricing on all CleanScribe RNA Polymerase products through June 30, 2026.
The 10% discount is already applied to the listed prices, no promo code required.
*Offer cannot be combined with other discounts, special offers, or existing price agreements.

Additional Resource

Read the technical bulletin where we highlight how CleanScribe RNA Polymerase:

  • Reduces dsRNA formation by up to 85% relative to the wild-type T7 RNA polymerase (mRNA or saRNA synthesis)
  • Exhibits comparable mRNA yield, capping efficiency, and integrity to WT T7 RNA polymerase
  • Further minimizes dsRNA in IVT reactions optimized for low dsRNA and high yield.

CleanScribe RNA Polymerase efficiently incorporates N1-methylpseudouridine. EGFP, FLuc, and Cas9 mRNAs containing CleanCap AG 3′ OMe or CleanCap M6 and N1-methylpseudouridine (replacing uridine) were synthesized using WT T7 or CleanScribe RNA polymerase and purified by LiCl precipitation. The resulting mRNAs were analyzed for (A) dsRNA levels by ELISA, (B) yield by UV spectrometry, and (C) capping efficiency by LC-MS.

CleanScribe RNA Polymerase offers a simple yet effective solution for dsRNA reduction during IVT.

  • Reduces dsRNA formation across various constructs, CleanCap cap analogs, and modified nucleotides
  • Directly replaces wild-type T7 RNA polymerase in diverse IVT protocols without compromising other critical quality attributes
  • Efficiently synthesizes long and complex templates like saRNAs while minimizing dsRNA formation
  • Improves mRNA performance by lowering innate inflammatory responses and increasing protein expression

The product may be ordered using the following catalog numbers.

Catalog number Total units Concentration Intented use
E-0107-S 5 000 U 50 000 U/mL Traditional protocols
E-0107-L 25 000 U
E-0107-001 5 000 U 100 000 U/mL TriLink protocols and specialized applications
E-0107-01 50 000 U
E-0107-08 420 000 U
E-0107-10 500 000 U 500 000 U/mL Protocol development

Unit definition: One unit of enzyme incorporates 1 nmol of ATP in 1 hour at 37°C.

Why Choose These 3D Culture Platforms?

  • Enhanced Physiological Relevance: Models that better mimic in vivo biology for disease research and therapeutic screening.

  • NAM-Aligned Methodologies: Support ethical innovation by reducing dependency on animal models and extending predictive in vitro capabilities.

  • Compatibility & Scalability: All systems fit standard lab infrastructure and integrate with imaging and automation workflows.

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  • References

    Article content created by Tebubio using courtesy materials provided by TriLink .