Custom Circular RNA (circRNA) Synthesis

Circular RNA (circRNA) is a single-stranded RNA molecule in which the 5' and 3' ends are covalently linked to form a closed loop. This circular structure offers several functional advantages, including increased stability and prolonged expression duration. CD Formulation specializes in high-quality custom synthesis of circRNA. Our customers need only to provide the desired sequence, and we offer a comprehensive service that encompasses everything from circRNA synthesis to analysis.

Background of Circular RNA (circRNA)

CircRNA is a single-stranded circular RNA produced through reverse splicing, often expressed by plasmids or viruses. However, the generation of circRNAs from plasmids or viruses involves auxiliary processes, such as reverse splicing, which may complicate the results of the study. By directly transfecting synthesized circRNAs, researchers can investigate the biological functions of circRNAs more effectively.

Fig.1 Features of Circular RNA. (CD Formulation)

Functions of Circular RNA (circRNA)

• Transcription Regulation

CircRNAs can bind to host genes at their synthesis sites, causing transcription pauses or termination by forming RNA-DNA hybrids (R-loop structures), which can lead to the skipping of exons or the production of truncated transcripts.

• Competing Endogenous RNAs

CircRNAs can regulate the expression of other genes by adsorbing microRNAs (miRNAs). This mechanism is akin to that of a CircRNAs absorb miRNAs, thereby diminishing the regulation of other target genes by these miRNAs.

• Synergistic Effects of Protein Interactions

CircRNAs can also bind to proteins, forming complexes that influence intracellular signaling and gene expression. These complexes may play crucial roles in the cell cycle, cell differentiation, and other biological processes.

• Coding Functions

Some circRNAs have the ability to be translated into proteins. Although this phenomenon is relatively rare, it adds additional functionality to circRNAs.

Explore Our Custom Circular RNA (circRNA) Synthesis Services

Chemical Synthesis of Cyclic RNA

The chemical in vitro synthesis of cyclic RNA primarily depends on phosphoramidite chemistry and solid-phase synthesis techniques. This process utilizes nucleoside triphosphate derivatives as raw materials and employs specific protecting groups to inhibit undesired chemical reactions that lead to the formation of 3'-5' phosphodiester bonds. This method enables precise control over the synthesis of RNA sequences and typically produces cyclic RNA comprising fewer than 70 to 80 nucleotides.

Enzymatic Ligation Synthesis of Cyclic RNA

Enzymatic ligation synthesis is performed through in vitro transcription (IVT), requiring a DNA template, reaction buffer, and phage RNA polymerase, primarily from T7, SP6, or T3 phages, with T7 being the most common.

General Workflow for Circular RNA (circRNA) Preparation

Fig.2 Flow chart of circRNA preparation. (CD Formulation)

• Sequence Design

We will select a cyclization strategy, design cyclization sites, and optimize codons to ensure that the RNA sequence exhibits strong expression and functional properties.

• Template Preparation

This step involves synthesizing a linear RNA template that will be utilized in the subsequent in vitro transcription (IVT) process, establishing the foundation for the generation of cyclized RNA.

• IVT-circRNA

The linear RNA template is transcribed into an RNA product with cyclization potential using in vitro transcription techniques, followed by the cyclization of the RNA with specific enzyme systems.

• Lipid Nanoparticle (LNP) Encapsulation

At this stage, the cyclized RNA is encapsulated in lipid nanoparticles (LNPs) to enhance its stability and delivery efficiency within the cell, thereby ensuring its effective function in biological systems.

Our Technology Platforms

Our Circular RNAs are prepared using both solid-phase synthesis technology and enzymatic synthesis technology. Additionally, they undergo characterization through a variety of analytical techniques to ensure their purity and validity. Below are the multiple analytical platforms we utilize.

Why Choose Us for Circular RNA (circRNA) Synthesis?

• We offer a comprehensive one-stop service that includes RNA synthesis and cyclization. Additionally, we accept linear RNA products provided by customers for subsequent cyclization.
• With state-of-the-art experimental equipment and exceptional technical support, we are committed to delivering the products our customers need in the shortest possible time.
• The specific cyclization-mediated sequence incorporated within the cyclization framework can effectively guarantee the accurate cyclization of the linear sequence of target circRNAs.

Publication Data

CD Formulation offers custom synthesis services for circular RNA (circRNA), emphasizing the delivery of high-quality circRNA molecules for research and biotechnology applications. Please feel free to contact us, and we will provide you with a reliable and efficient solution to support your research projects.

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.