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Custom circRNA Synthesis

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Custom circRNA Synthesis

Circular mRNA (circRNA) has been widely used in the study of gene or protein functions. Meanwhile, it is popular in the development of prophylactic and therapeutic biologics, similar to mRNA.

Yaohai Bio-Pharma has established a set of circRNA synthesis technologies to provide high-quality RNA, such as sequence design and optimization, in vitro transcription (IVT), circularization/cyclization, purification, and lipid nanoparticle (LNP) encapsulation. All products are released under the stringent quality control (QC) standards.

CircRNA Applications

Compared with linear mRNA, circular RNA (circRNA) is more stable in structure and is a major hot spot in current nucleic acid drug research. It has a wide range of applications in different fields, whether as coding circRNA or non-coding circRNA, including prophylactic vaccines for infectious diseases, therapeutic oncology vaccines, therapeutic oncology drugs, protein replacement therapy, regenerative medicine, cell and gene therapy (CGT), molecular sponge role and RNA aptamer.

Service Details
Process Optional Service Service Details Delivery Period (day)
circRNA Sequence Design and Optimization Design and Optimization of Coding Sequences CDS Codon Optimization 1
Design and Optimization of Non-Coding Sequences Intron and Exon Sequence, Homologous Arm and Spacer Design and Optimization 1-2
Circular Plasmid Preparation Gene Synthesis Gene Synthesis (Third-party) 7-10
Plasmid Amplification Plasmid Amplification 2
Plasmid Extraction
Linearized Plasmid Preparation Plasmid Linearization and Purification Plasmid Linearization 1
Linearization DNA Purification
In Vitro Transcription and Cyclization In Vitro Transcription and Cyclization In Vitro Transcription and Cyclization Reactions 1-2
RNase R Digestion
Optimization-optional Reaction Composition, Time Optimization 2-5
circRNA Purification Conventional Purification Method Lithium Chloride Precipitation 1
Magnetic Bead Purification 1
Highly Pure circRNA Purification Preparative High Performance Liquid Chromatography (HPLC) 1-2
Buffer Exchange Ultrafiltration 1
circRNA Lyophilization Sample Filling Filling 2-3
Lyophilization Pre-freezing
Primary Drying (Sublimation)
Secondary Drying (Desorption)
circRNA-LNP Encapsulation LNP Encapsulation Material and Liquid Pretreatment 2
Microfluidic Device Mixing
Buffer Exchange Tangential Flow Filtration 1
Sterilize Filtration
Quality Control of Plasmid DNA Concentration/purity Ultraviolet (UV) Spectrophotometry 1-2
Plasmid conformation Agarose Gel Electrophoresis (AGE)
Capillary Electrophoresis (CE)-Optional
Plasmid identify Restriction Enzyme Identification/AGE
Quality Control of circRNA Concentration/purity Ultraviolet (UV) Spectrophotometry -
Purity Agarose Gel Electrophoresis (AGE)/E-Gel 0.5
HPLC-optional 1
Quality Control of circRNA-LNP Encapsulation Efficiency RiboGreen Method 1
Particle Size Dynamic Light Scattering (DLS) 1
Polydispersity Index Dynamic Light Scattering (DLS) 1
Zeta Potential Dynamic Light Scattering (DLS) 1
Cell-based Potency Assay Cell Transfection Cell plating, cell transfection 4
Target Protein Detecting Fluorescence observation, Western Blot/ELISA 1-3
Case Study

We transfected enhanced green fluorescent protein circRNA (eGFP circRNA), mCherry circRNA, luciferase circRNA into 293T cells, and detected target protein through fluorescence, luminescent or color signal, western blot (WB), enzyme-linked immunosorbent assay (ELISA).

Cell-based Potency Assays of circRNA Samples by Yaohai Bio-Pharma

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