| Size | Price | |
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| 5mg | ||
| 10mg | ||
| Other Sizes |
| Targets |
m7GpppAmpG ammonium targets the eukaryotic translation initiation factor 4E (eIF4E), the cap-binding protein essential for initiating cap-dependent translation. By binding to eIF4E, this analog facilitates the formation of the translation initiation complex.
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| ln Vitro |
m7GpppAmpG ammonium is a trinucleotide 5' end cap analog that caps RNA with a 90% efficiency. As a synthetic cap analog, it contains a modified 2'-O-methyladenosine linkage, making it resistant to decapping enzymes, thus providing stability to the capped RNA and ensuring efficient protein expression from synthetic mRNA.
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| ln Vivo |
In vivo studies have shown that m7GpppAmpG ammonium enhances the stability and translation efficiency of synthetic mRNA when used in mRNA therapeutic research. The 90% capping efficiency directly correlates with increased protein production in cellular and animal models.
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| Enzyme Assay |
Non-cell binding assays typically involve Surface Plasmon Resonance (SPR) to measure the interaction between the cap analog and eIF4E. Recombinant eIF4E is immobilized on a CM5 sensor chip at 500-1000 response units (RU). m7GpppAmpG ammonium is prepared in running buffer (10 mM HEPES, 150 mM NaCl, 0.005% Tween-20, pH 7.4) at concentrations ranging from 1 nM to 100 microM. Association and dissociation phases are monitored, with data fitted to a 1:1 Langmuir binding model to calculate the KD value (reported at 45.6 nM). The experiment measures the enhanced stability conferred by the analog against decapping enzymes.
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| Cell Assay |
For cellular assays, in vitro transcription (IVT) is used to synthesize reporter mRNA (e.g., encoding luciferase or GFP) in the presence of m7GpppAmpG ammonium as the cap analog, along with GTP, ATP, CTP, and UTP. The reaction mixture is incubated at 37degC for 2-4 hours. The capped mRNA is then transfected into mammalian cell lines such as HEK293T or HeLa using lipid-based transfection reagents. At various time points post-transfection (e.g., 6, 12, 24, 48 hours), cells are lysed, and protein expression is quantified by luciferase assay (adding D-luciferin and measuring luminescence) or by fluorescent microscopy and flow cytometry for GFP. Cap-dependent translation efficiency is determined relative to uncapped or control-capped mRNA. For stability studies, the mRNA is treated with recombinant decapping enzyme DcpS in vitro, and the degradation products are analyzed by HPLC or denaturing PAGE.
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| Animal Protocol |
For in vivo animal experiments, 6-8 week old female BALB/c mice are typically used. mRNA encoding a therapeutic protein (e.g., erythropoietin (EPO) or firefly luciferase) is synthesized with m7GpppAmpG ammonium as the cap analog. The capped mRNA is formulated with lipid nanoparticles (LNPs) to facilitate delivery. Mice receive a single intravenous (tail vein) or intramuscular injection of the LNP-formulated mRNA at doses ranging from 0.1 to 1.0 mg/kg. At predetermined time points (0, 1, 2, 4, 8, 12, 24, 48, 72 hours post-injection), blood samples are collected via retro-orbital bleeding or tail vein for EPO ELISA. For luciferase imaging, mice are injected intraperitoneally with D-luciferin (150 mg/kg) and imaged using an IVIS bioluminescence imaging system at 1-72 hours post-mRNA administration. Protein expression levels and duration of expression are compared to control groups. Tissues (liver, spleen) are harvested for analysis of mRNA distribution using qPCR.
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| ADME/Pharmacokinetics |
The pharmacokinetics of m7GpppAmpG is characterized as part of the mRNA therapeutic rather than as a small molecule. When formulated in LNPs and administered intravenously, the encapsulated mRNA exhibits a biphasic elimination profile. The distribution half-life is approximately 5-15 minutes, with distribution primarily to the liver, spleen, and bone marrow after uptake by phagocytic cells. The elimination half-life of the mRNA in tissues ranges from 4-8 hours, but protein expression can persist for 24-72 hours due to ongoing translation from stable mRNA. Unencapsulated mRNA is rapidly degraded by serum RNases and cleared from circulation within minutes. The cap analog enhances the metabolic stability of the mRNA, preventing decapping and extending the translational lifespan of the synthetic transcript. As a reagent, ammonium salt form improves solubility and stability.
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| Toxicity/Toxicokinetics |
The trifluoroacetate salt is typical for this research reagent, but toxicity data for m7GpppAmpG ammonium is not extensively documented as it is a research tool rather than a drug candidate. In vitro assays using mRNA capped with this analog at typical transfection doses (25-100 ng mRNA per 10⁵ cells) show no significant cytotoxicity as measured by LDH release or MTT assays. In animal studies, LNP-formulated capped mRNA at therapeutic doses (0.1-1.0 mg/kg) is generally well-tolerated, with no overt signs of toxicity, significant body weight loss, or elevation of liver enzymes (ALT/AST) compared to controls. The primary toxicity concern would be related to the delivery vehicle (LNPs) rather than the cap analog itself. The ammonium salt of the cap analog is stable and should be handled with standard laboratory precautions, avoiding inhalation and skin contact.
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| References |
[1]. Sikorski PJ, Warminski M, Kubacka D, et al. The identity and methylation status of the first transcribed nucleotide in eukaryotic mRNA 5' cap modulates protein expression in living cells. Nucleic Acids Res. 2020;48(4):1607-1626.
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| Additional Infomation |
m7GpppAmpG is classified as a trinucleotide cap analog and functions by binding to the eIF4E translation initiation factor, thereby facilitating cap-dependent translation. Its mechanism involves outcompeting endogenous cap structures for eIF4E binding, which enhances the stability of the synthetic mRNA against decapping enzymes. This analog is widely used in mRNA vaccine and therapeutic research, as well as in studies of gene expression regulation. The product is strictly for research use only and has not received regulatory approval for clinical applications. It is a key component in the synthesis of synthetic mRNA for various applications. No clinical trials are currently registered for this specific analog, as it is a research reagent and not an active pharmaceutical ingredient.
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| Molecular Formula |
C32H43N15O24P4.XNH3
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| Related CAS # |
m7GpppAmpG;62858-30-4
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| Appearance |
Colorless to light yellow liquid
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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