T7 RNA Polymerase 3.0 GMP-grade (low dsRNA)
Low-dsRNA T7 RNA polymerase for IVT mRNA synthesis.
Product overview
During the in vitro transcription reaction, the by-product dsRNA generated can strongly stimulate intracellular RNA receptors and trigger innate immune signals. T7 RNA Polymerase 3.0 GMP-grade (low dsRNA), independently developed and multi-step purified by Biori Biotech to GMP standards, has high transcriptional catalytic activity for different templates and nucleic acid types. Compared with the common T7 RNA polymerase, T7 RNA Polymerase 3.0 (low dsRNA) significantly reduces the content of by-product dsRNA produced during transcription. T7 RNA Polymerase 3.0 (low dsRNA) can precisely recognize the T7 promoter region (5'-TAATACGACTCACTATAG-3') and start transcribing the subsequent DNA sequence into single-stranded RNA from the G in this region. Using natural or modified nucleotides as substrates, a single 20 μL in vitro transcription reaction can generate a large amount of RNA.
Available versions
| Cat. no. | Product name | Formulation |
|---|---|---|
| GMP-BP-E11C200-5K | T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) | |
| GMP-BP-E11C200-50K | T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) | |
| GMP-BP-E11C200-500K | T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) |
Features and applications
- 1 Effective control of dsRNA production.
- 2 Broad transcription compatibility: suitable for linearized templates of different lengths and for transcription with different nucleotide types.
- 3 High integrity of transcribed samples.
- 4 Sustained high transcription efficiency.
- 5 Strong transcriptional driving force: low dependence on Mg2+ during transcription.
- 6 The product has good stability.
- In vitro transcription and mRNA process development workflows.
- In vitro transcription with reduced dsRNA by-product formation, using natural or modified nucleotides as substrates.
Components
| Components | Cat. No. | Quantity | Volume |
|---|---|---|---|
| T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) | GMP-BP-E11-5K | 5 KU | 100 µL |
| GMP-BP-E11-50K | 50 KU | 1 mL | |
| GMP-BP-E11-500K | 500 KU | 10 mL |
Performance data
1 Testing the content of dsRNA in the transcription products
Description: The content of dsRNA in the transcription product was precisely quantified using ELISA, verifying the ability of T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) to control dsRNA during the transcription process.
Conclusion: dsRNA content analysis shows that, in the transcription of different lengths linearized templates, T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) produces low dsRNA content. The dsRNA level is reduced approximately 8-fold (7.9-fold and 8.5-fold for the 2000 nt and 5000 nt templates, respectively) compared with that produced by wild-type T7 RNA polymerase. Compared with T7 mutants from other suppliers, the product also shows an advantage in controlling dsRNA production.
2 Template compatibility test
Description: Verify the compatibility of T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) with templates of varying lengths and different nucleotide types during the transcription process.
Conclusion: The analysis of the yield indicates that, in transcription using linearized templates of different lengths and different nucleotide types, the yields of transcripts produced with T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) remain high.
3 Transcription product integrity test
Description: The integrity of the transcription products of T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) was verified using capillary electrophoresis.
Conclusion: Capillary electrophoresis data indicate that the transcriptional products of T7 RNA polymerase 3.0 exhibit high integrity.
4 Enzyme activity curve
Description: The T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) demonstrates sustained high efficiency across various transcription time points.
Conclusion: In the transcription of different length linearized templates, T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) maintains sustained high transcription efficiency.
5 Transcriptional driving force validation test
Description: Verify the dependence of T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) on Mg2+ in the transcription reaction buffer during the transcription process.
Conclusion: The experimental results show that Biori T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) retains strong driving capability during transcription at a relatively low working Mg2+ concentration (20 mM), outperforming the T7 RNA polymerases from Supplier A and Supplier C at this concentration. This indicates that T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) has low dependence on Mg2+.
6 Stability test
Description: After prolonged storage at 37 °C, the T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) was then verified for its stability through in vitro transcription.
Conclusion: After accelerated treatment at 37 °C for 7 days, T7 RNA Polymerase 3.0 GMP-grade (low dsRNA) still maintains a high transcription yield.
Shipping and storage
| Condition | Recommendation |
|---|---|
| Storage | Store at -20±5°C. |
| Shipping | Ship frozen on dry ice or with validated frozen cold-chain packaging that maintains the specified frozen storage condition. |
| Handling | Avoid repeated freeze-thaw cycles; mix thoroughly after thawing where applicable. |
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