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.

Features

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)

Components

Specifications

QC Standard

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

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.