Proprietary circRNA vector and purification method, leading to high circularization efficiency and purity
Improved stability & increased and longer protein expression duration vs linear mRNA
Lower immunogenicity vs Cap1 linear mRNA
Send Us Your ORF or IRES
We will design your circRNA expression vector with T7 promoter, PIE, IRES and ORF
Gene Synthesis to Plasmid Prep
Our integrated workflow will take care of all the prerequisites to produce your circRNA
RUO Grade IVT Production
µg to mg customized RUO circRNA service with advanced purification (RP-HPLC) and QC options
| QC | Item | Test Method | Specification | RUO Default |
RUO Upgrade |
|---|---|---|---|---|---|
| Identification | Appearance | Visual Inspect | Clear and free of foreign particles | ||
| RNA Length | Agarose Gel Electrophoresis | Expected size band detected | |||
| Identification | RNaseR degradation test | Not be degraded by RNaseR | |||
| RNA Content | UV Absorbance | Target ± 5% | |||
| PH | pH meter | Target ± 0.5 | |||
| Buffer Specification | Client Spec | N/A | |||
| Purity | A 260/280 Ratio | UV Spec | 1.70 ~ 2.30 | ||
| Purity by HPLC | HPLC | ≥80 % | |||
| Impurity | Total protein residue | Nano Orange Assay | ≤ 1% | ||
| Plasmid DNA Residue | qPCR | ≤ 0.1% | |||
| Safety | Endotoxin | Semiquantitative | < 10EU/mg | ||
| Endotoxin | Quantitative | < 10EU/mg | |||
| Bioburden | Sdirect Inoculation | No Growth after 48 hrs |
RNA Copy Number by RT-PCR
RNA Expression by Fluorescence Intensity
Circular eGFP- RNA (nonmodified) and Linear eGFP-mRNA (100% N1-me-pseU modified) were transfected with equal molar quantity in A549 cells. Coding RNA abundance was measured by qPCR, and GFP intensity was measured by flow cytometry. Results demonstrated that circRNA has higher stability, slower degradation, compared to linear IVT mRNA. circRNA also exhibits higher protein expression level and duration compared to linear IVT mRNA
GenScript circRNA demonstrates lower immunogenicity than mRNA. For IL6 test, ELISA method was used Abcam Human IL-6 ELISA Kit (ab46027). For INF-a and NF-kb test, A549 dual cells (InvivoGen) were transfected with test mRNA, and the expression of INF-a in cells was detected by relative luminescence testing for ISG54 activity following QUANTI-Luc/Gaussia kit instruction. The NF-Kb level was detected by OD at 630nm of cell medium following QUANTI-Blue kit instruction.
GenScript circRNA shows higher initial expression and cumulative expression than both mRNA and circular RNA prepared with a published method. A549 cells were transfected with circRNAs and mRNA encoding eGFP. circRNAs were prepared using GenScript’s proprietary method and using methods published in Nat Commun 9, 2629 (2018). Fluorescence was analyzed via flow cytometry for up to 6 days following transfection (10,000 cells were analyzed per condition at each time point). A549 cell doubling time = 22hr.
GenScript HPLC-purified circRNA shows higher expression than both crude circRNA and circRNA prepared with a published method. Cells were transfected with circRNAs encoding eGFP prepared using GenScript’s proprietary method and using methods published in Nat Commun 9, 2629 (2018). Fluorescence was analyzed via flow cytometry.
Circular RNAs (circRNAs) are a type of single-stranded RNA molecule with a covalently closed loop structure. They have been found to play important roles in various biological processes.
Some of the current applications of circRNAs include:
Overall, the study of circRNAs is an exciting and rapidly developing field, with many potential applications for improving human health.
Circular RNA introduction, GenScript offering details, and case studies showing increased stability, prolonged expression, and lower immunogenicity compared to linear IVT mRNA.
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High-quality custom mRNA production via our proprietary IVT RNA manufacturing workflow.
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Our off-the-shelf mRNA is optimized with 5’ cap and polyA tails that is ready to use for your research needs
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LNP Packaging
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Send us your in house mRNA samples for expert QC analysis.
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