The ScriptCap 2'-O-Methyltransferase Enzyme is derived from the Vaccinia virus and methylates the penultimate nucleotide in an eukaryotic mRNA transcript, converting a Cap 0 transcript into the natural Cap 1 mRNA structure. This methylation results in an up to 50% increase in the in vivo translation efficiency when compared to Cap 0 mRNA.
The enzyme has been shown to be active both on the natural Cap 0 structure and several different Cap 0 analogs used in co-transcriptional capping kits (See Table 1). Use of the enzyme is simple, as each reaction is capable of methylating 60 ug of Cap 0 RNA in 30 minutes.
When used in conjuction with the ScriptCap™ m7G Capping System, the ScriptCap 2'-O-Methyltranferase offers the only commercially available method to produce the Cap 1 structure.
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Table 1. Various different forms of capped and tailed mRNA were transfected into HeLa cells and assayed for luciferase activity. Data is normalized to the Cap 0, poly(A) transfection results. The complete Cap 1 mRNA exhibited up to 50% higher in vivo translation efficiency when compared to the various Cap 0-type structures currently used in most transfections.
| No RNA |
no |
none |
0% |
ScriptCap™ m7G Capping System
(Capping Enzyme) |
yes |
m7Gppp[m2'-O]N
(Cap 1) |
147% |
AmpliCap-Max™ High Yield
Message Maker Kit
(Standard Cap Analog) |
yes |
m7Gppp[m2'-O]N *
(Cap 1) |
148% |
AmpliScribe™ T7-Flash™ Transcription Kit
(ARCA Cap Analog) |
yes |
m27, 3'-OGppp[m2'-O]N *
(Cap 1) |
128% |
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