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YEASEN

[YEASEN] CleaScrip™ T7 RNA Polymerase (low dsRNA, 250 U/μL)

Cat-No. 10628ES

YEASEN은 생명과학 연구 및 진단을 위한 고품질의 연구용 시약과 기기를 제공하는 글로벌 바이오 기업입니다.

제품 설명

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

제품 번호

10628ES10 / size: 10KU

10628ES60 / size: 100KU

제품 설명

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
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	10628ES10	10628ES60	10628ES72	10628ES86
		(10 KU)	(100 KU)	(250 KU)	(2,500 KU)
10628	CleaScrip™ 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℃
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.