Vaccinia Capping Enzyme (High Efficiency, High Purity)

Description

Highly purified and extremely active Vaccinia Capping Enzyme (VCE)

For extremely efficient enzymatic formation of Cap0-capped mRNA from in vitro transcribed mRNA

Cap1 mRNA can be obtained by using VCE in combination with 2′-O-Methyltransferase (2OM)

• High purity-grade recombinant enzyme for mRNA manufacturing workflows involving post-transcriptional mRNA capping
• Integrates 3 enzymatic activities required for capping: RNA triphosphatase, guanylyltransferase, and guanine-N7-methyltransferase (requiring GTP, and SAM as a methyl donor)
• Efficiently adds an m⁷G cap to the 5′ end of IVT-generated mRNA producing Cap0-capped mRNA with up to 100% capping efficiency in a single reaction
  

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  The 5′ cap structure (m⁷GpppN) (Cap0), also known as the methylguanosine cap of mRNA is essential for translation initiation, protection from RNase degradation, and enhancement of mRNA stability during splicing and nuclear export.       Biori´s Vaccinia Capping Enzyme is a multi-step purified recombinant highly efficient enzyme preparation manufactured by Biori Biotech to GMP standards. The enzyme preparation integrates the three enzymatic activities required for capping: RNA triphosphatase, guanylyltransferase, and guanine-N7-methyltransferase. In the presence of S-adenosylmethionine (SAM) as a methyl donor as well as guanosine triphosphate (GTP), VCE directly and efficiently adds an m⁷G cap to the 5′ end of  in vitro transcribed mRNA in the correct orientation, producing Cap0-capped RNA with up to 100% capping efficiency in a single reaction. 

Contents:

1000 ul  Vaccinia Capping Enzyme (10 U/ul), 10000 Units, sufficient for 1000 Capping Reactions (based on the input of 10 U for 50 pmol denatured mRNA)

2000 ul  10x Capping Buffer

1000 ul    10mM GTP Solution

  200 ul    32mM SAM (S-adenosylmethionine)

NOTE:   Combining VCE with 2′-O-Methyltransferase (2OM) enables the formation of Cap1-capped mRNA from in vitro transcribed mRNA.

Cap1 (m7GpppNm), common in higher eukaryotes such as mammals, includes an additional methyl group at the 2′-O position of the first nucleotide, and has been shown to be important for evading the intracellular immune system, and to increase the stability and translational activity of the mRNA.