- Achieve ~100% Cap 0 or Cap 1 capping of IVT RNA, a level unattainable using cap analogs.
- Generate Cap 1-RNA in a single, 45-minute reaction.
- Enhance mRNA stability and translation efficiency through post-transcriptional 5’ capping.
- Directly input capped RNA into A-Plus™ Poly(A) Polymerase tailing reactions with no cleanup required.
Product is for research use only (RUO)
Product Description:
The ScriptCap™ Cap 1 Capping System provides both ScriptCap™ Capping Enzyme and ScriptCap™ 2'-O-Methyltransferase allowing users to produce either a Cap 0 or Cap 1 cap structure on the 5' end of in vitro transcribed
(IVT) RNA. ScriptCap™ Capping Enzyme catalyzes three enzymatic
reactions: (1) conversion of the 5′-triphosphate of RNA to a
diphosphate, (2) joining of GTP to the 5′-diphosphate of the first
nucleotide and (3) methylation of the 7-nitrogen of guanine using S-adenosyl-methionine
(SAM). These reactions produce Cap 0-RNA which can then be methylated
to Cap 1-RNA through the simultaneous use of ScriptCap™
2’-O-Methyltransferase. The system ensures nearly 100% 5’ capping
efficiency, a level that cannot be obtained using co-transcriptional
capping methods.
5’ Capping enhances mRNA stability and translation efficiency in
cells compared to uncapped RNA, with Cap 1 methylation further boosting in vivo translation
efficiency. A standard reaction caps 60 μg of RNA to Cap 0- or Cap
1-RNA in a single, 45-minute reaction and can be scaled up or down to
accommodate user needs.
ScriptCap™ capped RNA can be added directly to A-Plus™ Poly(A) Polymerase tailing reactions without cleanup for seamless synthesis of fully 5’ capped and 3’ poly(A) tailed mRNA.
Materials Supplied:
Important Store at -20°C in a freezer without a defrost cycle. Do not store at -70°C.
|
ScriptCap™ Cap 1 Capping System Contents |
|
Kit Component |
Reagent Volume |
C-SCCS1710 -
10 Reactions |
C-SCCS2250 -
50 Reactions |
|
ScriptCap™ Capping Enzyme, 10 U/μl |
40 μl |
200 μl |
|
ScriptCap™ 2'-O-Methyltransferase, 100 U/μl |
40 μl |
200 μl |
10X ScriptCap™ Capping Buffer
0.5 M Tris-HCl, pH 8.0, 60 mM KCl and 12.5 mM MgCl2 |
100 μl |
500 μl |
|
10 mM GTP |
100 μl |
500 μl |
|
20 mM S-adenosyl-methionine (SAM) |
50 μl |
250 μl |
|
ScriptGuard™ RNase Inhibitor, 40 U/μl |
25 μl |
125 μl |
|
RNase-Free Water |
0.67 ml |
3.35 ml |
Materials Required, but not Supplied
- IVT RNA
- Materials or kits for purification of the RNA product
REFERENCES
1. Kuge, H. et al., (1998) Nucl. Acids Res. 26, 3208.
2. Schlee, M. and Hartmann, G., (2016) Nat. Rev. Immunol. 16, 566.
3. Konarska, M.M. et al., (1984) Cell 38, 731.
4. Jemielity, J. et al., (2003) RNA 9, 1108.
5. Grudzien, E. et al., (2004) RNA 10, 1479.
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