Hieff™ T7 RNA polymerase (low dsRNA,250 U/μL)_ 10628ES yeasen
$180.00
In stock
SKU
10628ES10
Product Description
This is an engineered T7 RNA polymerase variant (low dsRNA) derived from the wild-type T7 RNA polymerase and produced in Escherichia coli. It significantly reduces the production of double-stranded RNA (dsRNA) while efficiently incorporating cap analogs, and exhibiting highly efficient in vitro transcription (IVT) comparable to the wild-type T7 RNA polymerase. It catalyzes the 5'→3' synthesis of RNA on double-stranded DNA from its T7 promoter sequence (5'-TAATACGACTCACTATAG*-3') and uses NTPs as substrates. Note: G* is the first base of the RNA transcript. The original brand name of the product is CleaScrip™.
Features
- Reduced dsRNA Content:
- dsRNA content is reduced by more than 10-fold.
- High Yield:
- Stable yield of over 9 mg/mL.
- Low Cap Analog Input**:
- Cap Analog input can be reduced to 2.5 mM.
- High Capping Efficiency**:
- Capping efficiency exceeds 99%.
- Please find information on the development of this enzyme.
Components
- Components No.
- Name
- 10628ES10
- 10628ES60
- 10628ES72
- 10628ES86
- (10 KU)
- (100 KU)
- (250 KU)
- (2,500 KU)
- 10628
- Hieff T7 RNA Polymerase (low dsRNA, 250 U/μL)
- 40 μL
- 400 μL
- 1 mL
- 10 mL
Specifications
| Source | Recombinant E. coli with T7 RNA Polymerase gene Optimum Temperature 37℃ |
| Endotoxin | <20 EU/mg Protease Negative Exonuclease Negative Nickase Negative RNase Negative E.coli host protein <50ppm E.coli host DNA <10 fg/U Mycoplasma examination Negative Related Products Tris NTPs synergistically decreasing dsRNA 10× Transcription Buffer 2 GMP-grade 10670ES N1-Me-Pseudo UTP Tris Solution GMP-grade (100 mM) 10657ES Pseudo UTP Tris Solution GMP-grade (100 mM) 10656ES ATP Tris Solution GMP-grade (100 mM) 10652ES GTP Tris Solution GMP-grade (100 mM) 10655ES CTP Tris Solution GMP-grade (100 mM) 10653ES UTP Tris Solution GMP-grade (100 mM) 10654ES Double-stranded RNA (dsRNA) ELISA kit 36717ES |
| Storage Buffer | 50 mM Tris-HCl, 1 mM EDTA, 10 mM DTT, 100 mM NaCl, 0.1% Triton X-100，50% (v/v) glycerin，pH7.9 at 25℃ |
| Unit Definition | The amount of enzyme required to incorporate 1 nmol of [ 3 H] GMP into the acid-insoluble precipitate within 1 hour at 37°C and pH 8.0 is defined as 1 unit. Recommended Mg2+ 30mM Magnesium Acetate QC Standard I tems Specification/Standard Enzyme |
| Activity | 250 300U/μL Protein Purity ≥95% |
Figures
1. 8K Sequence Validation – WT-T7 vs. T7 (Low dsRNA) T7 RNA Polymerase Yield(mg/mL) mRNA Integrity(%) dsRNA content (%) WT T7 9.5 81.9 0.29% Low dsRNA T7 9.0 82 0.0037% 2. Capping Efficiency & Cellular Function Validation: For both 4K and 1K fragments, capping efficiency was evaluated at different cap analog input amounts, showing that the input can be reduced to as low as 2.5 mM without significant differences in capping efficiency or cellular function at 48 h. No significant differences were observed between T7 mutant and wild-type (WT) enzymes. Customer feedback: E
Figure 1. Testing of T7 RNA Polymerase mutants. dsRNA Content: The dsRNA content reduced by more than 10-fold (A). mRNA Integrity: Maintained at over 90% (B). 2K Fragment Capping Efficiency: Exceeded 99% (C, D). High integrity (E) Table 1. The Yields of low dsRNA T7 RNA polymerase is comparable with that of WT.
Figure 2. Evaluation of the immunogenicity of the IVT products in murine RAW264.7 cells (Figures 2A and 2B). IFN-β mRNA and protein levels were reduced in RAW264.7 cells transfected with mRNA produced by mutants compared to wild-type, indicating that mRNA synthesized by the wild-type T7 RNAP elicited the strongest immune response, while mRNA from the mutants showed a significantly reduced response.
Figure 3. The dsRNA content in mRNA synthesized with Hieff™ T7 RNA polymerase is lower than that in mRNA synthesized with the wild-type enzyme after cellulose treatment.
Shipping and Storage
The products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year. Publication: Engineered T7 RNA polymerase reduces dsRNA formation by lowering terminal transferase and RNA-dependent RNA polymerase activities, The FEBS Journal 03 March, 2025 FADS and semi-rational design modified T7 RNA polymerase reduced dsRNA production, with lower terminal transferase and RDRP activities, bioxRxiv, 31 May, 2024
Safety Data Sheet
- 10628_MSDS_HB250724.pdf
Manuals
- 10628_Manual_Ver.EN20250724.pdf
Related Blog
- Hieff™ Low dsRNA T7 RNA Polymerase Mutants, Empowering mRNA Vaccine and Therapy Development
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