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Custom RNA Longmer Synthesis


RNA longmer has a wide range of applications in modern gene therapy technology, from understanding newly discovered genes to functional analyses of genes. These long RNA oligonucleotides can be used not only as a diagnostic tool for sensitive molecules, but also as a positive control for the development of medical therapy. Creative Biolabs uses DNA templates to provide customers with chemical synthesis services for long RNAs up to 120-300 bases in length. Our custom RNA longmer synthesis service includes custom mRNA, tRNA or other long non-coding RNA (lncRNA), and also provides RNA oligomerization services with various modifications and labels.

RNA Longmer Synthesis

As the oldest biological molecule, RNA molecules have rich functional applications in cells. With our deep understanding of its structure, more and more functionally specialized RNA molecules are synthesized and used in a variety of biological research. However, the length of 50-70 bases of RNA fragments in traditional synthesis methods has limited its use. Therefore, effective strategies for fragment ligation or other alternative novel approaches are needed.

Creative Biolabs' RNA synthesis service not only meets your needs for long-segment RNA synthesis, but also provides site-specific modification through multiple strategies.

Enzymatic Ligation of RNA Fragments

Ligation of synthetic RNA fragments is a basic method for preparing long modified RNAs. The required modifications can be incorporated into oligonucleotides of appropriate length by chemical synthesis, and then the individual fragments are ligated to form the full-length RNA. The most commonly used three different polynucleotide ligases are capable of ligating nicks in single- and / or double-stranded DNA or RNA constructs: T4 DNA ligase (T4 Dnl), T4 RNA ligase 1 (T4 Rnl 1) and T4 RNA ligase 2 (T4 Rnl 2). They have different requirements for the nicking state of the nucleic acid substrate, but they can be flexibly used according to the actual situation.

Ligation strategies involving ligases T4 Dnl, T4 Rnl 1, T4 Rnl 2. Figure 1. Ligation strategies involving ligases T4 Dnl, T4 Rnl 1, T4 Rnl 2. (Balke, 2014)

Chemical Ligation of RNA Fragments

Chemical ligation has attracted lots of attention from scientists as a method for preparing large fragment nucleic acids more than 20 years ago and has been continuously improved. From the initial use of compounds such as cyanogen bromide, water-soluble carbodiimide or 3-phosphate sulfate, to the emerging click chemical synthesis in recent years. We do this via a cycloaddition reaction between an azide and an alkyne-functionalized educt, the 1,3-dipolar Huisgen cycloaddition reaction. Triazole is formed in a buffered aqueous solution or an organic solvent, thereby forming 1,4- and 1,5-triazole regioisomers.

The Huisgen cycloaddition of an azide and an alkyne. Figure 2. The Huisgen cycloaddition of an azide and an alkyne. (Balke, 2014)

Internal Modification of Longmer RNA

In addition to the above-mentioned synthetic strategies, Creative Biolabs also carries out internal modification of RNA longmer for customers through precision-designed modification strategies, which can theoretically perform almost all types of site modification. Our service is based on small engineered binuclease applications that can mediate the exchange of short RNA patches within individually added synthetic RNA fragments in the appropriate RNA substrate.

Twin ribozymes for RNA functionalization. Figure 3. Twin ribozymes for RNA functionalization. (Balke, 2014)

Features

If you have any questions about our RNA longmer synthesis service, you can contact us by email or send us an inquiry to find a complete solution.

Reference

  1. Balke, D.; et al. (2014). Synthesis of Site-Specifically Modified Long-mer RNAs. Chemical Biology of Nucleic Acids. 477-496.

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