As the DNA template does not encode the 5' end cap, a synthetic cap analogue must be added to the mRNA. Cap analogs, which affect mRNA translation and transformation, are valuable tools for studying gene expression, and are often associated with therapy. Anti-reverse cap analog (ARCA) is a cap analog that produces immunogenic cap 0 for the production of capped transcripts during in vitro transcription.
As a platform technology, synthetic mRNA is increasingly used to express proteins needed in cell culture and even in vivo. The 5' cap of mRNA is an important feature that affects the stability and translation efficiency of mRNA, and ultimately affects the total protein content of mRNA expression. The capping of in vitro transcriptional mRNA is usually achieved by adding the corresponding dinucleotide cap analog to the reaction.
In the presence of cap dinucleotide, when the mRNA was blocked by transcription in vitro, it was found that m7GpppG cap analog could bind in two orientations, and only the correctly located derivatives, namely, m7G, were in the outer position and participated in the translation process. This has been solved by designing a modified cap analogue with O-CH3 replacing the 3'- or 2'- hydroxyl group of m7G, so it is called "ARCA".
Fig.1 Structural requirements for a cap analog to be incorporated into mRNA and to produce ARCAs.1
As a therapeutic drug for gene therapy, mRNA has huge application potential. It has been applied in some ongoing clinical trials, which is the driving force for the development of new mRNA cap modification molecular tools and the construction of new cap-based molecular probes. Synthetic cap analogs are useful research tools that can facilitate the study of cap-related processes at the molecular level and in biochemical and biological experiments, and may even eventually lead to the development of new therapeutic methods.
Due to the excellent inhibitory properties of ARCA, such compounds can help to prepare mRNA transcripts with high translation activity. To date, ARCAs cap RNA synthesized by in vitro transcription has been widely used in the study of mRNA function and metabolism, in vitro protein synthesis and expression of exogenous mRNA in living cells.
Creative Biolabs provides products and services with promising quality and exceptional price. Our mRNA service allows researchers to customize synthesis and introduce modifications to produce transcripts that are best suited for downstream applications. Based on years of intensive research, we provide customized high-quality ARCAs cap mRNA synthesis services. mRNA can be generated by DNA template provided by customers, or we can provide all-round services from scratch. Please contact us for more information.
Inquire About Our ServicesA: ARCAs caps ensure correct incorporation orientation, leading to higher translation efficiency and stability compared to traditional caps.
A: We use rigorous quality control measures, including analytical techniques to confirm cap structure and mRNA integrity.
A: Yes, we provide comprehensive consultation to tailor the ARCAs capping process to specific research needs.
A: Modifications include O-H, O-F, and O-S substitutions, which can alter biological activity and enhance protein binding.
A: By ensuring correct orientation, ARCAs caps significantly improve translation efficiency in vitro and in cultured cells.
The article investigates the use of anti-reverse cap analogs (ARCA) in enhancing the efficiency of mRNA transfection into mesenchymal stem cells (MSCs). The study explores the effects of different mRNA capping methods, including ARCA and its beta-thiophosphate derivatives (b-S-ARCA D1 and b-S-ARCA D2). It finds that b-S-ARCA D1 leads to higher protein production but with lower MSC metabolic activity, while ARCA ensures correct cap orientation, improving mRNA stability and translation efficiency. This research highlights the potential of ARCA-capped mRNA for clinical applications in cell-based therapies.
Fig.2 Metabolic activity of MSCs transfected with mRNA capped with different 5' cap analogs.2
| Cat. No | Product Name | Promoter |
|---|---|---|
| CAT#: GTVCR-WQ001MR | IVTScrip™ pT7-mRNA-EGFP Vector | T7 |
| CAT#: GTVCR-WQ002MR | IVTScrip™ pT7-VEE-mRNA-EGFP Vector | T7 |
| CAT#: GTVCR-WQ003MR | IVTScrip™ pT7-VEE-mRNA-FLuc Vector | T7 |
| CAT#: GTVCR-WQ87MR | IVTScrip™ pT7-VEE-mRNA-Anti-SELP, 42-89-glycoprotein Vector | T7 |
| Cat. No | Product Name | Type |
|---|---|---|
| CAT#: GTTS-WQ001MR) | IVTScrip™ mRNA-EGFP (Cap 1, 30 nt-poly(A)) | Reporter Gene |
| CAT#: GTTS-WK18036MR | IVTScrip™ mRNA-Human AIMP2, (Cap 1, Pseudo-UTP, 120 nt-poly(A)) | Enzyme mRNA |
| (CAT#: GTTS-WQ004MR) | IVTScrip™ mRNA-Fluc (Cap 1, 30 nt-poly(A)) | Reporter Gene |
| (CAT#: GTTS-WQ009MR) | IVTScrip™ mRNA-β gal (Cap 1, 30 nt-poly(A)) | Reporter Gene |
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