ARCA (Synonyms: Anti Reverse Cap Analog, 3´-O-Me-m7G(5')ppp(5')G) |
Catalog No.GC16608
One-Click Copy Product Info
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Anti-Reverse Cap Analog (ARCA) es un análogo de cap modificado en el cual el grupo 3' OH (más cercano a m7G) se reemplaza por –OCH3.
Products are for research use only. Not for human use. We do not sell to patients.
Sample solution is provided at 25 µL, 10mM.
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Anti-Reverse Cap Analog (ARCA) es un análogo de cap modificado en el cual el grupo 3' OH (más cercano a m7G) se reemplaza por –OCH3. Debido a esta sustitución, la ARN polimerasa solo puede iniciar la transcripción con el grupo hidroxilo restante, lo que obliga a la incorporación de ARCA en la orientación hacia adelante. Así, ARCA asegura la incorporación únicamente en la orientación hacia adelante. Como resultado, el 100% de los transcritos sintetizados con ARCA en el extremo 5' son traducibles, lo que lleva a un fuerte efecto estimulante en la traducción.
References:
[1].Janowski M, et al. The legacy of mRNA engineering: A lineup of pioneers for the Nobel Prize. Mol. Ther. Nucleic Acids. 2022 Sep 13; 29: 272-284.
Method for preparing correctly oriented cap-structured mRNA using ARCA[1]
This protocol is for reference only. Please adjust according to actual needs.
(1) Reagent preparation: dissolve ARCA in water or appropriate buffer to prepare a 0.5mM stock solution. Prepare an NTP mixture containing 0.5mM ATP, 0.5mM CTP, 0.5mM UTP and 0.1mM GTP (ARCA to GTP molar ratio 5:1). Prepare SP6 or T7 RNA polymerase, DNA template, RNase inhibitor and transcription buffer.
(2) In vitro transcription reaction system construction: add 600ng DNA template, 1U/μL RNase inhibitor, 0.5mM ATP/CTP/UTP, 0.1mM GTP and 0.5mM ARCA to the transcription buffer. Add SP6 or T7 RNA polymerase.
(3) mRNA synthesis: incubate at 37°C for 1 hour. Add 2μL DNase and incubate at 37°C for 20 minutes to remove template DNA. Purify the RNA product to obtain ARCA cap-structured mRNA with correct orientation.
(4) Translation verification of ARCA-capped mRNA: transfect ARCA-capped mRNA into HEK293 cells (DMEM+10%FBS; 37°C; 5%CO2), or add rabbit reticulocyte lysate and incubate at 30°C for 60 minutes. Evaluate translation efficiency by luciferase luminescence values. Detect remaining mRNA proportion in HEK293 cells by RT-qPCR to evaluate stability.
Precautions:
(1) To prevent RNA degradation, use RNase-free consumables throughout the experiment and perform operations on ice.
(2) For your safety and health, wear laboratory coats and disposable gloves during operation.
References:
[1] Kocmik I, Piecyk K, Rudzinska M, et al. Modified ARCA analogs providing enhanced translational properties of capped mRNAs. Cell Cycle. 2018;17(13):1624-1636.
| Cas No. | SDF | ||
| Sinónimos | Anti Reverse Cap Analog, 3´-O-Me-m7G(5')ppp(5')G | ||
| Canonical SMILES | OC1C(COP([O-])(OP([O-])(OP([O-])(OCC2OC(N3C=[N+](C)C4=C3N=C(N)NC4=O)C(O)C2OC)=O)=O)=O)OC(N5C=NC6=C5N=C(N)NC6=O)C1O | ||
| Formula | C22H32N10O18P3 (free acid) | M.Wt | 817.4 g/mol (free acid) |
| Solubility | Storage | Store at -20°C | |
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.2234 mL | 6.117 mL | 12.2339 mL |
| 5 mM | 244.7 μL | 1.2234 mL | 2.4468 mL |
| 10 mM | 122.3 μL | 611.7 μL | 1.2234 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A liquid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 31 reference(s) in Google Scholar.)