Vaccinia Capping System
Based on the Vaccinia Virus Capping Enzyme, the Vaccinia Capping System (NEB# M2080) provides the necessary components to add 7-methylguanylate cap structures (Cap 0) to the 5´ end of RNA. In eukaryotes, these terminal cap structures are involved in stabilization, transport and translation of mRNAs. Enzymatic production of capped RNA is an easy way to improve the stability and translational competence of RNA used for in vitro translation, transfection and microinjection. Alternatively, use of labeled GTP in a reaction provides a convenient way to label any RNA containing a 5′ terminal triphosphate.
Advantages of the Vaccinia Capping System
- Capping mRNA prior to in vivo or in vitro translation
- Labeling 5′ end of mRNA
RNA Cap Analogs
The 5′ terminal m7G cap present on most eukaryotic mRNAs promotes in vitro and in vivo translation, at the initiation level. For most RNAs, the cap structure increases stability, decreases susceptibility to exonuclease degradation, and promotes the formation of mRNA initiation complexes. Certain prokaryotic mRNAs with 5′ terminal cap structures are translated as efficiently as eukaryotic mRNAs in a eukaryotic cell-free protein synthesizing system. Splicing of certain eukaryotic substrate RNAs has also been observed to require a cap structure.