Custom Circular RNA Synthesis

Circular RNA (circRNA) has gained prominence as a crucial component of the transcriptome, highlighting its essential role in gene expression regulation and cellular function. It is characterized by its covalently closed loop structure, which offers a level of stability and functionality not found in traditional RNA forms. As the landscape of RNA biology evolves, circRNAs are increasingly recognized for their potential applications in diagnostics, therapeutics, and biotechnological innovations. At BOC Sciences, we are committed to facilitating the exploration and application of circRNA through our specialized Custom Circular RNA Service, tailored to meet the diverse needs of researchers and institutions.

What is Circular RNA?

Circular RNAs are a distinct class of non-coding RNAs that form a closed-loop structure due to a unique splicing mechanism known as back-splicing. This process occurs when a downstream splice donor connects with an upstream splice acceptor, resulting in a continuous RNA molecule devoid of free ends. This characteristic not only enhances the stability of circRNAs but also contributes to their resilience against degradation by cellular exonucleases. CircRNAs were first discovered in viruses, but subsequent studies have revealed their widespread presence in eukaryotic organisms, including plants and animals. High-throughput sequencing technologies have unveiled thousands of circRNAs in human cells alone, underscoring their abundance and biological significance. Furthermore, circRNAs can arise from exons, introns, or both, leading to diverse isoforms with varying functions.

What is the Advantage of Circular RNA?

The advantages of circRNA stem from their structural and functional attributes, making them particularly appealing in both basic research and applied sciences.

• Enhanced Stability: CircRNAs' closed-loop structure provides remarkable stability compared to linear RNAs. This resistance to degradation allows circRNAs to persist longer within the cellular environment, potentially leading to prolonged biological effects.
• Translational Potential: Some circRNAs have been identified as having open reading frames (ORFs) that enable them to be translated into peptides. This capability broadens their functional roles beyond conventional non-coding RNAs and positions them as potential sources of novel proteins.
• Tissue-Specific Expression: Many circRNAs exhibit distinct expression patterns across different tissues and developmental stages. This specificity suggests that they may have important roles in tissue homeostasis, development, and response to environmental stimuli.
• Role in Disease: The dysregulation of circRNA expression has been associated with various diseases, including cancer and neurodegenerative disorders. This link positions circRNAs as potential biomarkers and therapeutic targets.

Circular RNA vs Linear RNA

The distinctions between circular and linear RNA are profound, influencing their respective roles in cellular processes and applications in research.

• Structural Integrity: The most notable difference is the closed-loop structure of circRNAs, which renders them more stable and less susceptible to exonuclease activity compared to linear RNAs, which have exposed ends that are prone to degradation.
• Biogenesis Pathways: CircRNAs are synthesized through back-splicing, a process that differs from the canonical splicing of linear mRNAs. This unique biogenesis pathway contributes to the diversity of circRNA isoforms, which can include exonic, intronic, and hybrid forms.
• Functional Roles: While linear RNAs, especially mRNAs, serve primarily as templates for protein synthesis, circRNAs are involved in a variety of functions, including acting as miRNA sponges, participating in transcriptional regulation, and engaging with RNA-binding proteins.
• Research and Therapeutic Applications: The inherent stability and multifunctionality of circRNAs position them as attractive candidates for therapeutic applications, biomarker discovery, and innovative research tools, contrasting with the traditional roles of linear RNAs.
• Gene Regulation: CircRNAs can influence gene expression at multiple levels, acting not only as sponges for miRNAs but also by modulating transcription and translation processes, thereby participating in a complex regulatory network.

BOC Sciences' Custom Circular RNA Service

At BOC Sciences, we offer a comprehensive Custom Circular RNA Service designed to facilitate the synthesis and analysis of high-quality circRNA for various research applications. Our Custom Circular RNA Service provides researchers with access to high-quality circRNA tailored to their specific experimental needs. This service encompasses the entire process, from design and synthesis to characterization and quality control.

Our Service Workflow

• Consultation and Project Scoping: We initiate the service with a thorough consultation to ascertain the specific goals and requirements of the client. This includes discussing the desired circRNA sequences, modifications, and intended applications.
• Design and Bioinformatics Analysis: Utilizing advanced bioinformatics tools, our team designs optimal circRNA sequences, considering factors such as potential back-splicing efficiency, secondary structure predictions, and interactions with miRNAs and RNA binding proteins.
• Synthesis of Circular RNA: Employing state-of-the-art synthesis techniques, we produce high-yield circRNA. Our synthesis process is meticulously optimized to ensure the integrity and quality of the final product.
• Comprehensive Characterization: Each batch of synthesized circRNA undergoes rigorous quality control assessments. This includes verifying structural integrity using techniques such as gel electrophoresis, assessing yield and purity, and functional validation to confirm biological activity.
• Documentation and Delivery: The final product is delivered along with comprehensive documentation, including detailed analysis reports, sequence information, and recommendations for experimental applications.

Advantage of BOC Sciences' Custom Circular RNA Service

• Customized Solutions: Our service is designed to accommodate the unique needs of each client, providing circRNA products that are specifically tailored for their research goals and experimental designs.
• Expertise and Experience: With over two decades of experience in RNA biology and molecular genetics, BOC Sciences offers unparalleled expertise in the synthesis and application of circRNA.
• Commitment to Quality: We prioritize the quality of our circRNA products through stringent quality control measures, ensuring that clients receive reliable and reproducible results for their research.
• Comprehensive Support: Our dedicated team provides ongoing support throughout the project, assisting clients with experimental design, data interpretation, and guidance on circRNA applications in their research.
• Rapid Turnaround Times: We understand the urgency often associated with research projects. Our efficient workflow ensures that clients receive their custom circRNA promptly without compromising quality.

Applications of Circular RNA

The applications of circular RNA are diverse and continually expanding, reflecting their potential in various fields of research and biotechnology.

• Biomarker Discovery: CircRNAs' unique stability and tissue-specific expression profiles make them promising candidates for developing biomarkers for early disease detection, particularly in cancers and other chronic diseases.
• Therapeutic Development: CircRNAs are being explored for their potential as therapeutic agents in gene therapy, where they may be used to modulate gene expression or deliver therapeutic RNA molecules.
• Vaccine Research: The innovative properties of circRNAs have positioned them as potential candidates for vaccine development, particularly in the context of mRNA vaccines. Their stability and immunogenicity are currently under investigation.
• Regenerative Medicine: CircRNAs are being studied for their roles in stem cell biology and tissue regeneration, where they may influence cell fate decisions and promote tissue repair mechanisms.
• Functional Genomics: Researchers utilize circRNAs as tools in functional genomics studies to investigate gene regulatory networks, cellular signaling pathways, and mechanisms underlying various diseases.

BOC Sciences is dedicated to advancing circRNA research through our Custom Circular RNA Service, empowering scientists with high-quality, tailored solutions to explore the myriad applications of circRNA in their respective fields. As research into circRNA continues to unfold, its implications for diagnostics, therapeutics, and our understanding of fundamental biological processes promise to be profound and transformative.

Case Study

Case Study 1 Harnessing Circular RNA as a Novel Vaccine Platform for Potent T Cell Activation and Antigen Presentation

This case study examines the efficacy of circular RNA (circRNA) as a dual-function vaccine platform that acts both as an immunogen and adjuvant, highlighting its ability to induce potent T cell responses. Researchers investigated the immune response generated by circRNA encoding antigenic proteins, specifically through a charge-altering releasable transporter (CART). The study demonstrated that circRNA vaccinations, administered via various routes (subcutaneous, intranasal, and intravenous), resulted in robust antigen-specific CD8 T cell activation and strong antibody production. Notably, the circRNA-based vaccine outperformed traditional adjuvants, such as AddaVax and Poly(I), by facilitating significant innate immune activation and enhancing memory responses. This innovative approach not only provides insights into the immunogenic potential of circRNAs but also paves the way for their application in developing effective vaccines against infectious diseases and cancer therapies, suggesting a promising future for circRNA in RNA medicine.

FAQ

1. What is the difference between circular RNA and mRNA?

Circular RNA (circRNA) differs from mRNA in its structure, stability, and function. While mRNA has a linear structure with distinct 5' and 3' ends, circRNA forms a closed loop, providing enhanced stability and resistance to degradation. Furthermore, circRNAs primarily function as regulators of gene expression and cellular processes, whereas mRNA serves as a template for protein synthesis.

2. What viruses have circular RNA?

Several viruses are known to produce circular RNA, for example:

• Hepevirus: The hepatitis E virus has been shown to generate circRNAs during its replication cycle.
• Caronaviruses: Certain coronaviruses, such as SARS-CoV-2, can produce circular RNA as part of their life cycle.

These viral circRNAs have implications in host-pathogen interactions and may contribute to the pathogenesis of viral infections.

3. How to design primer for circular RNA?

Designing primers for circRNA amplification involves several key considerations:

• Target Site: Select a region within the circRNA that is specific and avoids linear RNA contamination.
• Back-Splicing Junction: Primers should flank the back-splicing junction of the circRNA to ensure specificity.
• Melting Temperature (Tm): Ensure primers have similar Tm values for optimal annealing during PCR.

4. How to identify circRNA?

Identifying circRNAs involves a combination of experimental techniques, including:

• RNA Sequencing (RNA-Seq) profiles RNA transcripts, including circRNAs. RNA is isolated, treated with exonucleases to enrich for circRNA, and sequenced using next-generation sequencing (NGS).
• Polymerase Chain Reaction (PCR) amplifies specific circRNAs using primers targeting back-splicing junctions. After reverse transcription to synthesize complementary DNA (cDNA), PCR products are analyzed via gel electrophoresis to detect circRNA.
• Northern Blotting allows for circRNA detection by separating total RNA via gel electrophoresis, followed by hybridization with labeled probes specific to circRNA.
• Fluorescence In Situ Hybridization (FISH) visualizes circRNA within cells. Specific probes are designed for the target circRNA, and after hybridization, fluorescence microscopy is used to observe circRNA localization.

5. Can ecDNA be transcribed into circular RNA?

Extrachromosomal DNA (ecDNA) can indeed be transcribed into circular RNA. Recent studies have demonstrated that ecDNA is a source of circRNA, particularly in cancer cells. The presence of ecDNA-derived circRNAs may contribute to tumorigenesis and provide insights into the molecular underpinnings of cancer.

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Custom Circular RNA Synthesis

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