Next-Generation T7 RNA Polymerase

Unlocking the Power of Next-Generation T7 RNA Polymerase: A Guide to the T7 RNAP Variant Toolbox

Explore GENTAUR’s T7 RNA Polymerase Variant Toolbox featuring T7-D13, T7-C10, and T7 Turbo. Learn how these next-gen enzymes improve mRNA synthesis, reduce dsRNA contaminants, and enhance transcription integrity in synthetic biology.

Introduction to T7 RNA Polymerase in Synthetic Biology

T7 RNA polymerase (T7 RNAP) has become the backbone of in vitro transcription (IVT) in modern synthetic biology and mRNA production. However, conventional T7 RNAP often presents challenges, such as double-stranded RNA (dsRNA) contamination, poor capping efficiency, and instability at elevated temperatures. These issues can compromise mRNA therapeutic quality, translational efficiency, and safety.

To address these limitations, GENTAUR has introduced a toolbox of advanced T7 RNAP variants — T7-D13, T7-C10, T7-P02, T7-M1, and T7 Turbo — each engineered to resolve specific bottlenecks in high-yield and high-integrity RNA synthesis workflows.

                                                                                                 

GENTAUR's T7 RNAP Toolbox : Key Variants and Their Advantages

T7-D13: The State-of-the-Art dsRNA Reducer

T7-D13 stands out for its ability to reduce dsRNA by ~100-fold compared to wild-type (WT) T7 RNAP. With residual dsRNA levels < 2 ppm, this variant ensures a significantly cleaner mRNA output — a critical quality attribute for therapeutic RNA.

-Peer-reviewed validation: He W, et al. “Effective Synthesis of High-Integrity mRNA Using In Vitro Transcription.” Molecules 29.11(2024).

T7-C10: Capping Cost Reduction Without Compromise

Capping is a major cost driver in mRNA synthesis, especially with cap analogs such as Cap 1. T7-C10 innovates by boosting cap analog binding affinity (KdCap/KdNTP), allowing :

  • 95% reduction in cap analog input
  • Continued use of wild-type GG promoters

Results across various mRNA sequences confirm ~95% capping efficiency, maintaining comparable integrity to WT.

-Peer-reviewed validation : He W, et al. “Effective Synthesis of mRNA during In Vitro Transcription with Fewer Impurities Produced.” Molecules 29.19(2024):4713.

Performance Comparison: T7 RNAP Variants vs. Wild-Type

Variant Yield (mg/mL) Integrity (%) Capping Efficiency (%) Residual dsRNA (%)
T7-D13 8.3 97.8 97.9 0.0080
T7-C10 8.4 94.1 95.2 0.0127
WT T7 8.1 93.2 94.3 0.019