mRNA Cap 2'-O-Methyltransferase GMP-grade

Methyltransferase for converting Cap0 RNA to Cap1 RNA.

Cap0 → Cap1

High-efficiency 2′-O-methylation

mRNA Cap 2'-O-Methyltransferase GMP-grade, GMP-BP-E06

Product overview

2'-O-methyltransferase (2OM) is a methyltransferase encoded by the vaccinia virus DNA and expressed in recombinant E. coli. It is a methyltransferase that further modifies the Cap0 cap structure into the Cap1 cap structure. Using S-adenosylmethionine (SAM) as the methyl donor, this enzyme adds a methyl group to the 2'-O position of the first nucleotide immediately adjacent to the Cap0 cap structure at the 5' end of the RNA, thereby generating an mRNA bearing the Cap1 structure. This modification can further reduce the intrinsic immunogenicity of the mRNA and enhance the expression level of the encoded protein post-transfection.

Available versions

Cat. no.	Product name	Formulation
GMP-BP-E06-5K	mRNA Cap 2'-O-Methyltransferase GMP-grade
GMP-BP-E06-50K	mRNA Cap 2'-O-Methyltransferase GMP-grade
GMP-BP-E06-500K	mRNA Cap 2'-O-Methyltransferase GMP-grade

Features and applications

Features

Recombinant 2'-O-methyltransferase expressed in E. coli; activity: 50 U/μL.
Unit definition: one unit is defined as the amount of enzyme required to incorporate 10 pmol of methyl groups into Cap0 RNA in 1 hour at 37°C.
Quality control: protein purity ≥95%; free of exogenous DNase, RNase, exonuclease, and endonuclease activity; residual host-cell DNA ≤100 pg/mg; residual host-cell protein <50 ppm; bacterial endotoxin <5 EU/mL.

Applications

Conversion of Cap0 RNA to Cap1 RNA in a stepwise capping system after Vaccinia Capping Enzyme capping.
One-step Cap1 capping in combination with Vaccinia Capping Enzyme GMP-grade (GMP-BP-E05).

Components

Components	GMP-BP-E06-5K	GMP-BP-E06-50K	GMP-BP-E06-500K
	5 KU	50 KU	500 KU
mRNA Cap 2'-O-Methyltransferase GMP-grade (50 U/µL)	0.1 mL	1 mL	10 mL
10× Capping Buffer	0.2 mL	2 mL	20 mL
SAM (32 mM)	20 µL	0.2 mL	2 mL

Performance data

1 Schematic of Cap0 to Cap1 conversion.

Description: Schematic illustration of the enzymatic conversion from Cap0 to Cap1, where methylation of the 2'-hydroxyl group on the first nucleotide of the Cap0 structure generates the Cap1 structure (m⁷GpppNm).

GMP-BP-E06 mRNA Cap 2'-O-Methyltransferase GMP-grade: Figure 1 Principle scheme of Cap0 to Cap1 conversion. — **Figure 1** Principle scheme of Cap0 to Cap1 conversion.

Conclusion: Illustration of the enzymatic process converting Cap0 to Cap1, in which methylation of the first ribose 2'-hydroxyl group of Cap0 produces Cap1 (m⁷GpppNm).

2 High translational efficiency

Description: Uncapped GFP mRNA was capped with Vaccinia Capping Enzyme GMP-grade (VCE, GMP-BP-E05) alone to obtain Cap0 mRNA, or with VCE together with 2'-O-methyltransferase to obtain Cap1 mRNA. Upon cell transfection, the mRNA exhibited high expression levels, resulting in strong fluorescence signals.

GMP-BP-E06 mRNA Cap 2'-O-Methyltransferase GMP-grade: Figure 2 The fluorescence intensity results of mRNA-GFP prepared at different stages. — **Figure 2** The fluorescence intensity results of mRNA-GFP prepared at different stages (uncapped; Cap0 after VCE capping; Cap1 after capping with VCE and 2OM).

GMP-BP-E06 mRNA Cap 2'-O-Methyltransferase GMP-grade: Figure 3 Statistical results of the fluorescence intensities of mRNA-GFP prepared at different stages. — **Figure 3** Statistical results of the fluorescence intensities of mRNA-GFP prepared at different stages (GFP: uncapped; GFP-Cap0: VCE; GFP-Cap1: VCE and 2OM).

Conclusion: Cap1 mRNA prepared using Biori's 2'-O-methyltransferase together with VCE showed higher fluorescence intensity after transfection than Cap0 mRNA capped with VCE alone.

Shipping and storage

Condition	Recommendation
Storage	Store at -20±5°C.
Shipping	Ship frozen on dry ice or with validated frozen cold-chain packaging that maintains the specified frozen storage condition.
Handling	Avoid repeated freeze-thaw cycles; mix thoroughly after thawing where applicable.