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Cat. No.: OSMCK-20 (for 20T)

Cat. No.: OSMCK-100 (for 100T)

Description

One-step mRNA Capping Kit is a specialized reagent kit designed to efficiently add a Cap-1 structure to mRNA in a single-step reaction. It contains Faustovirus Capping Enzyme (FCE), mRNA Cap 2'-O-Methyltransferase (mRNA 2'-O-MTase), GTP, S-adenosylmethionine (SAM), and the necessary reaction buffer, ensuring streamlined and effective capping.

Faustovirus Capping Enzyme (FCE) is a single-subunit mRNA capping enzyme derived from the Faustovirus S17 strain through expression and purification. It possesses the three essential enzymatic activities required to add a Cap-0 structure to mRNA: Triphosphatase (TPase), Guanylyltransferase (GTase), and (Guanine-N7)-Methyltransferase (N7 MTase). These activities enable FCE to catalyze the addition of an N7-methylguanosine (m7GpppNp) cap at the 5' end of eukaryotic mRNA, thereby generating Cap-0 mRNA. This cap structure enhances mRNA stability by protecting it from 5'→3' exonuclease degradation and prevents triphosphorylated mRNA from triggering the innate immune response.

mRNA Cap 2'-O-Methyltransferase utilizes S-adenosylmethionine (SAM) as a methyl donor to add a methyl group at the 2'-O position of the first nucleotide adjacent to the Cap-0 (m7GpppNp) structure, resulting in Cap-1 (m7GpppNmp) mRNA. The Cap-1 modification further enhances mRNA stability and translation efficiency, improving expression levels following microinjection or transfection. Additionally, Cap-1 helps mRNA evade innate immune responses in certain cells.

Components

• Faustovirus Capping Enzyme
• mRNA Cap 2'-O-Methyltransferase
• Capping Buffer (10X)
• RNase Inhibitor (40U/μl)
• GTP (10mM)
• SAM (32mM)
• Ultrapure Water

Applications

• Capping mRNA for in vivo or in vitro translation.
• 5' end labeling of mRNA.
• 2'-O methylation in the Cap-1 structure facilitates immune evasion.
• Improved translation efficiency of mRNA.
• Enhanced protein expression after microinjection or transfection.

Storage

Store at -20ºC for long-term stability, with a minimum shelf life of two years.

Precautions

• This product should be kept in an icebox or on ice during use and stored at -20ºC immediately after use.
• Since RNA-related procedures are involved, strict adherence to RNA handling protocols is required to prevent RNase contamination.
• Pipette tips, centrifuge tubes, and other laboratory consumables must be RNase-free or treated with DEPC.
• Appropriate handling methods should be selected based on the specific application. Additional reagents, such as RNase Inhibitor or ultrapure water, may be required.
• SAM is unstable at pH 7-8 and 37ºC. It is recommended to prepare fresh working solutions based on the actual number of reactions just before starting the experiment. Keep the solution on ice to prevent degradation.
• This product is for scientific research use only by professionals. It must not be used for clinical diagnosis or treatment, food or drug applications, or stored in residential settings.
• For your safety and health, wear a lab coat and disposable gloves while handling this product.

Related:

• One-step mRNA Capping Kit
• mRNA Cap 2'- O-Methyltransferase
• Faustovirus Capping Enzyme (FCE)

SBS Genetech is recognized as one of the global major leading industry players in mRNA Raw Material by third-party market researchers. For more details, please visit Top 5 mRNA Vaccine & Therapeutics Raw Material Market Companies in Global Market 2022.

Only for research and not intended for treatment of humans or animals

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