RNA is a ubiquitous molecule that is found in most living organisms and viruses. It consists of nucleotides, where ribose sugars are bonded to nitrogenous bases and phosphate groups. The nitrogenous bases in RNA contain adenine, guanine, uracil, and cytosine. RNA is mainly single-stranded, while a few specific RNA viruses possess a double-stranded structure. The RNA molecule displays diverse lengths and structures. RNA is the genetic material for viruses instead of DNA and leads to many human diseases. Transcription refers to the conversion of DNA into RNA, while translation involves the synthesis of proteins based on the information encoded in RNA. The process of RNA synthesis and its functions are different between eukaryotes and prokaryotes. Some RNA molecules play a role in regulating gene expression and have the potential to be utilized as therapeutic agents in the treatment of human diseases. Therefore, it is significant to synthesize those molecules.
Fig.1 The structure of RNA. (Wikipedia, 2023)
CD BioGlyco provides two approaches to effectively synthesize RNA, including chemical synthesis and enzymatic synthesis. Our techniques are widely applicable and highly preferred for conducting structural studies on RNA molecules. We also offer a wide variety of Modifications for the RNA we synthesize.
Chemical synthesis of RNA
We utilize solid-phase chemical synthesis to acquire RNA by using an automated synthesizer. The method relies on the iterative elongation of the RNA chain on a solid support such as controlled-pore glass or highly cross-linked polystyrene in a cyclic manner.
Phosphoramidites decorated with different protection groups are employed to prevent their reactivity during chemical synthesis.
We offer a variety of phosphoramidites with a gentle base-labile protection of the exocyclic amino group of nucleobases and a phosphate moiety blocked by 2-cyanoethyl N, N, N', N'-tetra-isopropyl.
We utilize [(triisopropylsilyl)oxy]methyl (TOM) and test-butyldimethylsilyl (TBDMS) as the primary protection groups for the 2'-hydroxyl group.
Fluoride-labeled groups are eliminated by using tetra-n-butylammonium fluoride (TBAF) or triethylamine trihydrofluoride (TEA·3HF).
We typically use dimethoxy trityl (DMT) to block the 5' OH group, which is compatible with the 2' OH silyl blockage.
In our standard solid-phase synthesis, the elongation process takes place from the 3' to the 5'-end. Each elongation cycle adds one nucleotide, resulting in a gradual extension of the chain. During each cycle, there are four essential steps involved: deacetylation, coupling, capping, and oxidation.
Fig.2 Steps in solid-phase synthesis. (CD BioGlyco)
Enzymatic synthesis of RNA
We also provide an in vitro transcription method for synthesizing RNA molecules. This method involves the use of a DNA template containing a T7 RNA polymerase promoter sequence, followed by a sequence that encodes the target molecule. The T7 RNA polymerase enzyme binds to the promoter region of the DNA template and initiates RNA synthesis. The elongation of the RNA chain ceases when the enzyme dissociates from the 3'-end of the template. Our in vitro synthesis method enables the effective production of RNA, typically in the milligram range.
CD BioGlyco has a wealth of expertise in the synthesis of RNA. We have advanced technology and a skilled research team to meet your needs, whether it's targeted synthesis for specific sequences or the production of modified RNA molecules. We are committed to delivering high-quality custom RNA synthesis to help you in various research fields. If you have inquiries or are interested in our services, please feel free to contact us without hesitation.
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