CleaScrip™ T7 RNA Polymerase (GMP-Grade, low dsRNA, 250 U/μL) _ 10629ES
CleaScrip™ T7 RNA Polymerase (GMP-Grade, low dsRNA, 250 U/μL) _ 10629ES
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CleaScrip™ T7 RNA Polymerase (GMP-Grade, low dsRNA, 250 U/μL) _ 10629ES CleaScrip™ T7 RNA Polymerase (GMP-Grade, low dsRNA, 250 U/μL) _ 10629ES

CleaScrip™ T7 RNA Polymerase (GMP-Grade, low dsRNA, 250 U/μL) _ 10629ES

SKU: 10629ES10
Size: 10 KU
Price:
$198.00

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In stock
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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.

Feature

  • dsRNA level lower to about 1/100000
  • Comptiable with Trilink CleanCap AG, LZCap
  • High yields comparable to WT 
  • Lower Cap input
  • Animal origin-free (AOF)

Please find information on the development of this enzyme.

Components

Components No.

Name

10629ES10

10629ES60

10629ES72

10629ES86

 

 

(10 KU)

(100 KU)

(250 KU)

(2,500 KU)

10629

CleaScrip™ T7 RNA Polymerase (GMP Grrade, 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℃

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

Customer feedback:
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).
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 CleaScript™ 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:

FADS and semi-rational design modified T7 RNA polymerase reduced dsRNA production, with lower terminal transferase and RDRP activities

Documents

 

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    FAQ

    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.

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