DNA-Free mRNA-Capping Enzyme

The vaccinia virus capping system uses vaccinia virus capping enzyme and other components to add a 7-methylguanosine cap structure (Cap 0) to the 5´ end of RNA. In eukaryotes, the 7-methylguanosine cap structure affects mRNA stability, transport, and translation. Enzymatically capping RNA is a simple and effective method to enhance the stability and translation of RNA used in in vitro transcription, transfection, and microinjection. Additionally, labeled GTP used in the reaction provides a convenient labeling method for any RNA with a 5´ triphosphate end.

Cat. No.: DFMCE-400 (for 400U)

Cat. No.: DFMCE-2k (for 2000U)

Description

The vaccinia virus capping system uses vaccinia virus capping enzyme and other components to add a 7-methylguanosine cap structure (Cap 0) to the 5´ end of RNA. In eukaryotes, the 7-methylguanosine cap structure affects mRNA stability, transport, and translation. Enzymatically capping RNA is a simple and effective method to enhance the stability and translation of RNA used in in vitro transcription, transfection, and microinjection. Additionally, labeled GTP used in the reaction provides a convenient labeling method for any RNA with a 5´ triphosphate end.

The vaccinia virus capping enzyme consists of two subunits (D1 and D12). The D1 subunit, which is the mRNA-capping enzyme catalytic subunit, has RNA triphosphatase, guanylyltransferase, and guanine methyltransferase activities. The D12 subunit, which is the mRNA-capping enzyme regulatory subunit, regulates the capping activity of the D1 subunit. These activities are necessary to add a complete Cap 0 structure, i.e., m7Gppp(5´)N. The capping reaction is not only highly efficient, achieving up to 100%, but also ensures correct orientation, unlike co-transcriptional capping which may add some cap analogs.

Our DNA-Free enzymes are all recombinant proteins and purified through multiple steps, devoid of genomic and plasmid DNA from the production strain.

Unit Definition

One unit is defined as the amount of enzyme required to incorporate 10 pmol GTP into one 80-nucleotide transcript within 60 minutes at 37℃ under 1× mRNA-capping reaction buffer conditions.

Features

• Completely free from bacterial DNA.
• Capping of mRNA for in vitro or in vivo translation.
• Labeling the 5´ end of mRNA.