mRNA and saRNA capping
The 5' cap protects RNA against premature degradation and is required for translation initiation. Cap structure is also essential for distinguishing from non-self RNAs by the innate immune response. Our mRNAs and saRNAs are available with enzymatic Cap1 or with ExPLoRNA Therapeutics cap analogs (i.e. ExploCap™).
Cap1 or ExploCap™ RNAs bear biological properties equivalent to CleanCap® (a registered trademark of TriLink Biotechnologies). Capping efficiencies are above 90% in all cases.
| ARCA | CAP1 | ExploCap™ | CleanCap® | |
|---|---|---|---|---|
| Capping efficiency | 60-80% | 95-99% | 95-99% | 95-99% |
| Transcription Yield | Low | High | High | High |
Messenger RNA (mRNAs)
OZ Biosciences offers mRNAs that mimic fully processed mature mRNAs. These mRNAs are stabilized with 5’ Cap1 (enzymatic) or ExploCap™ (co-transcriptional) structure and 3’ poly(A) tail and are optimized to yield improved stability & performance. They are available unmodified or with nucleotide modifications that reduce innate immune responses: 5-methoxyuridine (5moU, replacing uridine) or N1-methylpseudouridine (m1Ψ).
Ready-to-use mRNA range:
- Reporter gene mRNAs: ideal controls to study transfection efficiency: eGFP, FLuc, RLuc, mCherry, β-Gal & Tomato mRNA .
- Fluorescent mRNAs: eGFP and FLuc mRNA fluorescently labeled with Cy5, Cy3, or AlexaFluor488.
- Genome editing mRNAs: Cas9, CRE and Cas13d mRNA.
- Vaccine mRNAs: ideal as controls for immunization or vaccine studies: SARS-CoV-2 spike (Delta, Omicron), influenza HA (H1N1, H3N2) and OVA.
- Gene Replacement mRNAs: EPO mRNA.
- Cytokines & Interleukin mRNAs: h IL-2, h IL-6, h CCL2, TGFα1, TNFα, TGFβ1, IFNα & IFNγ mRNA.
- Transcription Factor mRNAs: c-Myc, h Sox2, Nanog, KLF4, LIN28 and OCT4 mRNA.
- Cancer mRNAs: p95HER2, TP53, PD-L1 mRNA.
Self-amplifying RNA (saRNAs)
OZ Biosciences offers saRNAs able to express the gene of interest up to 1 month after transfection. They are designed to produce high expression level of protein with enhanced length of duration. These saRNAs are stabilized with 5’ Cap 1 structure and 3’ poly(A) tail. They are either unmodified, or modified with 5-methyl-Cytidine (m5C replaces C) to reduce innate immune responses.
Ready-to-use saRNA range:
- Reporter Gene saRNAs: eGFP and Fluc saRNA
- Vaccine/Antigen saRNAs: SARS-CoV-2 spike, influenza HA (H1N1) and OVA.
Circular RNA (circRNAs)
OZ Biosciences offers circRNAs, covalently closed RNA rings produced by in vitro transcription followed by ribozyme-mediated circularization. They are unmodified, protected from exonuclease-mediated degradation and translated through an internal ribosome entry site (IRES) instead of a 5' cap.
Ready-to-use circRNA range:
- Reporter Gene circRNAs: controls to study transfection efficiency: eGFP circRNA.
Custom mRNA and saRNA Synthesis Service
With over 20 years of nucleic acid delivery expertise, OZ Biosciences is confident in providing you with the best-in-class service for the production of high quality mRNAs & saRNAs.
We are able to synthesize custom messenger RNA (mRNAs) and self-amplifying RNA (saRNAs) at microgram to multigram scales, from a few hundred up to several thousand bases, with a wide variety of modifications including 5’ terminal modifications with cap, internal modification such as 5-methoxyuridine (other modified nucleotides are available) and 3’ modification such as poly-A tail. Fluorescent labeling with Cy5, Cy3 or other options is available.
OZ Biosciences' RNA production process has been meticulously perfected over the years to achieve an exceptional level of optimization. Using state-of-the-art enzymes, considered to be the most efficient available, the process guarantees production that minimizes dsRNA formation. Production follows four steps, adapted to the material provided:
Gene Synthesis
Depending on the nature of the sequence you provide (nucleotide or peptide sequence, PCR product or plasmid), this step includes gene synthesis, cloning, sequence verification and plasmid DNA linearization.
In Vitro Transcription (IVT)
We synthesize RNA by in vitro transcription (IVT) and offer a wide variety of co- or post-transcriptional modifications: capping, modified nucleotides, fluorescent labeling and poly(A) tail.
Downstream Process
The RNA is purified and supplied in the buffer and at the concentration of your choice, with a standard format of 1 mg/mL in citrate buffer, pH 6.4.
Quality Control
We assess the quality of the RNA through a series of analyses: size and integrity, concentration, purity and sterility. Additional analyses are available on request.
We ship your fully custom-made RNA in 2 to 6 weeks (depending on the source of the DNA template).
Quality control
We do know that the reproducibility of scientific experiments is very important to researchers. That is why we developed a quality control system tailored to each product, enabling us to detect defective batches immediately.
Our mRNAs & saRNAs, whether custom or catalog, undergo standard quality control, including measurement of size, integrity and purity by fragment analyzer, agarose gel and Nanodrop (concentration 260/280 and 260/230), a sterility test and an experimental validation (transfection / expression).
However, we have developed additional tests to extend the analysis to include endotoxin detection and double strand RNA (dsRNA) quantification.
| Items | Specification | Standard QC | Superior Grade QC* |
|---|---|---|---|
| Integrity | Agarose gel mobility and HPLC | ✓ | ✓ |
| Concentration | 1mg/ml +/- 5% | ✓ | ✓ |
| A260/280 |
>1.8 for Unmodified mRNAs >1.7 for chemically modified mRNAs |
✓ | ✓ |
| Sterility | Absence of bacterial growth at 37°C | ✓ | ✓ |
| Functionality** | Test for protein expression | ✓ | ✓ |
| Endotoxin | <0.5 EU/mL | ✓ | |
| dsRNA | <0.05% | ✓ |
*Our catalogue mRNAs undergo the standard QC. Superior Grade QC can be performed as an additional prestation. ** For reporter mRNAs and Spike SARS-Cov2 related mRNAs only for catalog mRNAs. Can be included in superior Grade QC.
Example of Quality Control results :
New enzyme for low dsRNA in our mRNAs!
As part of our continuous quality improvement, we have selected a new generation of RNA polymerase that reduces dsRNA during IVT. We are able to reduce the amount of dsRNA in our catalog and in our customized mRNAs by up to 88%.
Our new quality control limit is 0.05% of dsRNA.
Comparison of dsRNA amount in different mRNA produced by OZ Biosciences before and after the production process improvement.
Technical Resources
Want to discuss your research needs?
Our expert product support team is at your service for your special requests.
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