mRNA Cap 2´-O-Methyltransferase adds a methyl group at the 2´-O position of the first nucleotide adjacent to the cap structure at the 5´ end of the RNA.
mRNA Cap 2´-O-Methyltransferase adds a methyl group at the 2´-O position of the first nucleotide adjacent to the cap structure at the 5´ end of the RNA. The enzyme utilizes S-adenosylmethionine (SAM) as a methyl donor to methylate capped RNA (cap-0) resulting in a cap-1 structure.
Recently, it has been reported that the 2′-O-methylation in the cap-1 structure helps the mRNA evade innate immune response in some cell types in vivo (1,2).
mRNA Cap 2´-O-Methyltransferase specifically requires RNA with an m7GpppN cap as substrate. It cannot utilize RNA with pN, ppN, pppN or GpppN at the 5´ end (3,4). Capped RNA may be prepared via in vitro transcription using cap analog or through enzymatic capping using the Vaccinia Capping Enzyme (NEB #M2080 ).
The reagents provided in this pack can be used to methylate up to 400 μg of capped RNA.
The following reagents are supplied with this product:
NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
1X Capping Buffer
Supplement with 0.2 mM S-adenosylmethionine (SAM)
Incubate at 37°C
1X Capping Buffer
50 mM Tris-HCl
5 mM KCl
1 mM MgCl2
1 mM DTT
(pH 8 @ 25°C)
20 mM Tris-HCl
100 mM NaCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1% (w/v) Triton® X-100
pH 8 @ 25°C
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