Hieff™ T7 RNA polymerase (low dsRNA,250 U/μL)_ 10628ES
Hieff™ T7 RNA polymerase (low dsRNA,250 U/μL)_ 10628ES
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Hieff™ T7 RNA polymerase (low dsRNA,250 U/μL)_ 10628ES Hieff™ T7 RNA polymerase (low dsRNA,250 U/μL)_ 10628ES
Hieff™ T7 RNA polymerase (low dsRNA,250 U/μL)_ 10628ES
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...

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Hieff™ T7 RNA polymerase (low dsRNA,250 U/μL)_ 10628ES

YeasenSKU: 10628ES10
Size: 10 KU
Price:
$180.00
Stock:
In stock

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™.

Feature

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
(10 KU)

10628ES60
(100 KU)

10628ES72
(250 KU)

10628ES86
(2,500 KU)

10628

Hieff T7 RNA Polymerase (low dsRNA, 250 U/μL)

40 μL

400 μL

1 mL

10 mL
[Note]: T7 RNA Polymerase can be used in combination with 10× Transcription Buffer 3 (Cat#10674).

Specifications

Source

Recombinant E. coli with T7 RNA Polymerase gene

Optimum Temperature

37℃

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 [3H] 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

Items

Specification/Standard

Enzyme activity

250-300U/μL

Protein Purity

≥95%

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

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:
Figure 1. Testing of T7 RNA Polymerase mutants. dsRNA Content: The dsRNA content reduced by more than 10-fold (A).&nbsp; mRNA Integrity: Maintained at over 90% (B).&nbsp; 2K Fragment Capping Efficiency: Exceeded 99% (C, D). High integrity (E)
E
Figure 1. Testing of T7 RNA Polymerase mutants. dsRNA Content: The dsRNA content reduced by more than 10-fold (A).&nbsp; mRNA Integrity: Maintained at over 90% (B).&nbsp; 2K Fragment Capping Efficiency: Exceeded 99% (C, D). High integrity (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 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.

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.

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

Storage

The products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

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The product is for research purposes only and is not intended for therapeutic or diagnostic use in humans or animals. Products and content are protected by patents, trademarks, and copyrights owned by Yeasen Biotechnology. Trademark symbols indicate the country of origin, not necessarily registration in all regions.

Certain applications may require additional third-party intellectual property rights.

Yeasen is dedicated to ethical science, believing our research should address critical questions while ensuring safety and ethical standards.