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RiboPro is a full-service provider of custom and off-the-shelf messenger RNA (mRNA) and long-non-coding RNA (lncRNA). Our service contains 4 key elements, that you may order in combination or as stand-alone service.
We design mRNA with the desired expression profile (high-low, long-short, conditional) by exchanging UTRs and/or optimizing the codon usage in the coding sequence.
Particular attention is given to de-immunization of the mRNA, which can be achieved equally well with sequence engineering, as with modified nucleotides.
Effects of mRNA design
mRNA design influences expression a lot; the 5'UTR, the 3'UTR, the codon usage, secondary structure, type of capping and A-tail length all influence performance (total amount of protein expression) as well expression kinetics (expression duration, peak- vs - steady-state expression, etc.).
Sequence design by RiboPro
By choosing for RiboPro, you get access to mRNA experts that can turn any wild-type coding sequence into a high-performance mRNA. They will either design the complete mRNA, or just the parts (e.g. the UTRs) you request.
When designing your mRNA, our scientists use proprietary software to test in silico every combination of tried and tested UTRs and coding sequences designed with our patent pending de-immunization method until they find an mRNA that fits your application.
We use highly innovative, proprietary methods to synthesize high-quality, high-purity full-length mRNA.
Our enzyme-based methods produce mRNA virtually devoid of dsRNA, with high capping efficiency and well-defined poly(A)-tails, from either de novo or customer delivered DNA templates. If needed, we can arrange de novo DNA template synthesis.
High purity full-length RNA synthesis
RiboPro has developed a proprietary system using in-house developed enzymes with enhanced stability, activity and fidelity combined with progressively optimized protocols to ensure efficient synthesis of pure, full-length mRNA. These protocols allows for the production of short (around 100nt) to extremely long (15k nt) RNA transcripts with a minimum of side-products and contaminants. Any residual impurities are removed by our triple purification system, reducing such impurities at least 1000-fold.
To turn the basic RNA transcript into a fully functional mRNA, capping (addition of a 5' cap) and A-tailing is required. We offer 2 methods of capping; with our enzymatic Vaccinia capping system >99% capped mRNA can be achieved for uncomplicated 5'UTRs, whereas complicated 5'UTRs (with major or very stable secondary structure) can be capped co-transcriptionally with ARCA (Anti-reverse cap analog) or another cap analog. RiboPro has developed a proprietary method to ensure >95% capping with ARCA (100% correct orientation), where traditional protocols provide 50-80% capping via ARCA.
For the poly(A)-tail, we offer an enzymatic method producing a narrow distribution of fully-customizable A-tail lengths, and a template-based method producing a highly defined A-tail length. Frequently requested lengths (120/150nt) can be provided from stock templates.
Any mRNA purchased from RiboPro can be formulated into a selection of nano-particle technologies.
These technologies include cationic peptides, cationic or ionizable polymers and lipids. Our specialty is formulation in lipid nanoparticles (LNPs) of which we have several standard formulations available that display high mRNA delivery efficiency.
Our formulation service includes:
You may choose from various options:
Not everyone has access to a lab or the time to do basic mRNA testing. Or, you may lack mRNA-specific knowledge or equipment. When ordering mRNA from RiboPro, you don't have to have any of that; you have access to RiboPro's CRO services.
All our CRO services are executed by experienced scientists.
RiboPro offers the following mRNA-focused (standard) CRO services.
Other tests/assays may be available upon discussion. Contact our RiboProfessionals to learn what we can do for you!
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If you choose to use our coding sequence de-immunization, and/or a high number of other modifications, and/or if you do not have a suitable DNA template available, de novo DNA synthesis may be the fastest and most affordable way to go. In each of these cases, we would be happy to arrange de novo DNA synthesis as part of our service. Once we have the DNA template on file, next orders of the same sequence can be delivered in 1 week and at reduced cost.
If you do have a suitable template available, you may send us 1ug of high purity DNA for each 200ug of RNA you order. Once the DNA arrives, we perform several quality checks, linearize the DNA and start RNA synthesis.
The finished product is aliquoted in 1 or more screw cap tubes, depending on preference. We typically recommend aliquoting in multiple tubes as a countermeasure against RNAse-contamination. The tubes are individually labelled and allow visual inspection of the content.
Because of the sensitive nature of (m)RNA molecules, we ship on dry ice (-80oC), which is the golden standard for (m)RNA storage. We pack our products so that they survive at least 5 days at ambient temperature. Therefore, we only ship on Mondays to avoid materials being thawed over the weekend. Our ultra-pure RNA is stable for multiple days at 4oC and for weeks at -20oC, so alternative shipping solutions are available upon request.
You may choose from various options:
As scientists ourselves, we understand the pressures you are under, and time is of the essence. Therefore, we optimized our process to finish mRNA synthesis in 2-3 days.
If you send us a ready-to-use template arriving Wednesday latest, you may expect your product to be send the next Monday.
If your order requires de novo DNA synthesis, it typically takes 2-3 weeks (incidentally up to 4 weeks due to COVID-19 related busyness at our DNA synthesis partner) to complete your order. Please be aware that certain sequences present difficulties, due to secondary structure formation, repeats and other sequence features, and may require revised synthesis conditions, resulting in some delay. Of course, we always update you if and when this happens.
Because mRNA is a sensitive molecule, we ship on dry ice and are therefore bound to shipping on Mondays.
At RiboPro, we are committed to making your life easier. Thus, we are working hard on also making the ordering of mRNA quick and easy. At the moment, you may order in the following ways:
Next, we get to work: we check your sequences, assess the work to be done and provide you with a quotation within 24h (mostly within 2 hours, so monitor your mailbox).
If you agree with the quotation, we get started immediately with at-risk production while you arrange a P.O. number, or we wait for the P.O. number to be arranged (based on your preference).
During production, we do the following things:
For planning purposes, you will receive updates at steps 2, 4 and 6.