Custom mRNA

Built on more than 10 years of experience in mRNA synthesis, we are your trust-worthy source of custom messenger RNA. From µg-scale up to 100 grams, and lengths up to 15knt, we have you covered.

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  • Highest quality mRNA
  • Fastest turn-around
  • Support for success
  • Sensible scales

Custom mRNA synthesis

Our custom mRNA service produces high quality mRNA based on custom sequence designs, which may originate from various sources:

  1. your own sequence design, manufactured exactly according to your specification
  2. your sequence, optimized with our technologies for optimal expression
  3. a sequence designed by us, based on the protein you like to express.

Our fully enzymatic process is able to use several types of DNA as input, including:

  1. circular DNA plasmid provided by you or your DNA supplier
  2. circular DNA plasmid cloned by RIBOPRO
  3. linear DNA from an de novo DNA synthesis, and/or PCR method

 

Choices

When ordering custom mRNA you have various choices and options to customize the product to your exact needs. Is your favorite option not listed? Please contact us and we are happy to discuss a custom project with you.

Choice Description
Capping Co-transcriptional (ARCA)
Enzymatic (VCE/FCE)
Capping efficiency 80-95% (ARCA)
95-99% (VCE/FCE)
Cap-type Cap-0 (for standard cell-types)
Cap-1 (for immune cells & in vivo applications)
Poly(A)tail length 0-300nt
Segmented
Amount 100 µg – 100 g
Buffer PBS (pH 7.4)
Citrate (pH 5.5)
Other buffer

How to get a quote?

Tells us via our Quotation form exactly what you want and you will receive an initial quotation as soon as possible. In the form you can also add questions, remarks and ask our RiboProfessionals™ to contact you to help make adjustments to your mRNA-design/mRNA-project. Filling out the form is fast, easy and prepares our people to support you the best they can!

Get a quote Schedule a meeting

What will happen next?

After your quotation/order request

Once you send the form in, we take the following steps a.s.a.p. to help you get the best mRNA as fast as possible:
1. a RiboProfessional™ checks your request,
2. gets in touch if needed, and
3. sends you a quotation within 24h
4. when you OK the quotation, we get started.

For repeat orders (an option in our ordering menu), you will still get a quotation, but we often already start with synthesis ‘at risk’ from DNA that is ‘on file’ to realize the shortest possible turn-around time. Of course, you may still ask for amendments of the order based on the quotation. The risk is entirely ours.

After your order

During production, we do the following things:

  1. Optimize the sequence if desired
  2. Communicate optimized sequences back to you (optional)
  3. Order DNA if needed
  4. Perform RNA synthesis
  5. Perform Quality Control
  6. Arrange shipping
  7. Check if material has arrived complete and correct
  8. Send invoice
  9. Follow-up to help you with your experiments if needed

For planning purposes, you will receive updates at steps 2, 4 and 6.

FAQ

Answers to some of your burning questions:

  • Why not synthesize mRNA yourself?

    Of course as a life-science researcher you have the knowledge and tools to produce any bio-molecule yourself. However, several issues make this difficult for (m)RNA.

    The disadvantages of self-synthesis of (m)RNA

    So also for the synthesis of mRNA; the multi-step enzyme-based reaction, with 1 or more purifications, in addition to acquiring a suitable DNA template, can be time-consuming; taking 2-3 days per synthesis.

    In addition, especially when a limited amount or number of mRNAs is required, the up-front cost for the reaction ingredients, the quality control, the certified RNAse-free buffers and consumables is high.

    But perhaps more than the reasons listed above, very few labs are well-equipped to work with such a delicate molecule, which is subject to degradation to omni-present and hard-to-destroy RNAses. Working with mRNA as a final product in an experimental setting is certainly doable, but multi-step manipulations during synthesis in a sub-optimal environment present a significant risk to the integrity of the final product.

    And finally, designing a high-performance mRNA requires very specific knowledge on 5’UTR and 3’UTR functioning, secondary structure formation of RNA, the induction of innate immunity by RNA, codon optimization, ideal A-tail length, etc.

    Why choose RiboPro custom mRNA synthesis instead?

    By ordering your desired mRNA from RiboPro’s custom mRNA synthesis service you can get started with your experiment as quickly as possible, without hassle! No need to worry about RNAse contamination during production, no need to setup complex quality control, no need to invest in the equipment and materials; we take care of it all. Our RNA experts make sure you get the best results.

  • What is included in the service?

    RiboPro’s custom mRNA service is very flexible; we include as much or as little as you need.

    Generally speaking, we have 2 flavors:
    A. You provide us with a (semi)suitable template, which we use as input for the RNA synthesis. Minor changes to the template to make it suitable are no problem for us.
    B. You provide us with the desired sequence, and we perform de novo DNA synthesis to obtain a suitable template for the RNA synthesis.

    What is included?

    So, in the full-service variant (B) the following is included:
    1. mRNA design and optional sequence optimization
    2. de novo DNA synthesis
    3. DNA prep
    4. RNA synthesis
    5. RNA capping (up to 99% capped, and Cap1 modification possible)
    6. A-tailing to desired length (up to 500nt)
    7. 2 purification round
    8. (optional) dsRNA removal procedure
    9. Quality control (length, sequence verification (DNA), A-tail length, purity)
    10. (optional) Aliquotation
    11. Packaging & shipping
    12. After-care to ensure success in your experiments

    This very complete package is available from €660 + shipment cost (typ. €50-€150) + DNA synthesis (typ. €100-€200)

  • How does RIBOPRO optimize mRNA sequences?

    The performance of a messenger RNA, to be added to cells via standard transfection methods, greatly depends on the design of the sequence, the quality of the synthesis, chemical modifications of specific bases and the purity of the end-product. Introducing a sub-optimal mRNA into cells is known to trigger the innate immune response; resulting in lower protein expression, cellular toxicity and the release of pro-inflammatory cytokines.

    High performance mRNA
    For the best performance, we recommend the following combination of features to be selected for your custom mRNA:

    • 5’UTR: no upstream ATG, kozak or similar present directly upstream of the start codon. For example RiboPro catalogue 5’UTRs.
    • 3’UTR: selected from mRNAs known to be extra-ordinary stable.
    • Cap1 – 95%.
    • 120-150nt A-tail.
    • dsRNA reduction.

    These parameters will provide you with a decent expressing mRNA. However, for any of the following situations:
    • therapeutic applications,
    • a demand for high protein-expression,
    • experiments incompatible with pro-inflammatory cytokine release,
    • or any other application requiring the best protein expression and the lowest toxicity,
    we recommend additional mRNA-specific sequence-optimization. Using RiboPro’s patent-pending technology allows for 5x more protein (as high as 40x more in specific applications) and significant reduction in pro-inflammatory cytokine production.

    If you like to understand why these settings lead to the best performance, read more below.

    The trouble avoided by RiboPro’s high-quality RNA synthesis
    Synthesizing a high-performance messenger RNA requires optimization all through-out the multi-step synthesis, including specialized purification.

    dsRNA
    First of all, during the enzymatic production process, dsRNA can be formed. dsRNA is one of the main triggers of innate immunity, primarily via TLR3 (endosomal recognition during transfection), OAS1, PKR and MDA5 (cytoplasmic sensors). RiboPro’s mRNA synthesis process results in the lowest amount of dsRNA possible. The method used to reduce the amount of dsRNA, also negatively affects the yield of ssRNA, so limited additional costs do apply compared to the standard method, because we increase the scale in order to obtain the guaranteed yield. Without dsRNA reduction, protein expression is typically 10-fold lower than with dsRNA reduction.

    Uncapped, triphosphorylated RNA
    Secondly, currently no supplier can promise that 100% of all mRNA molecules will be capped. When uncapped, the 5’UTR of the mRNA contains a 5′ triphosphate, which can be a ligand for the cytoplasmic innate immune sensor RIG-I. RiboPro offers 2 capping grades (~80% and 95%+). If selecting the lower grade (cheaper), additional phosphatase treatment may be required to reduce RIG-I activation.

    Cap methylation status
    Third, if a cap is present, but no additional methylation of the first nucleotide (Cap0), then IFIT1 may be activated in certain cells, leading to around 30-50% reduction in activity in some experiments. This seems to be particularly relevant in cells of the immune system. RiboPro offers optional methylation of the first nucleotide (Cap1).

    Contaminants
    Finally, template DNA, salts and small RNA species (formed during abortive cycling of the RNA polymerase) must be carefully removed to avoid interfering with uptake and interpretation of results. RiboPro uses extensive DNAse I-based DNA removal, reducing the residual DNA to far less than 1 millionth of the RNA concentration.

    QC
    Next to an optimized production process, RiboPro also has also developed a series of test for quality control, offering you peace of mind about the identity, purity and modification status of your order.

    Do you have additional questions? Please contact us directly below or later via the contact page. We are happy to help!

  • How fast can we deliver?

    For a typical sequence, you will receive your order within 4-5 weeks. If you provide us with a suitable template, it may even be faster, because 3rd party de novo DNA synthesis is the most time-consuming step of our RNA synthesis process.

Custom fluorescent mRNA synthesis

If you need a special mRNA sequence with fluorescent-labelling, or if you need a special label itself, then you may order custom mRNA with a pre-defined degree of mRNA labelling.

Our custom mRNA service produces high quality mRNA based on custom sequence designs, which may originate from various sources:

  1. your own sequence design, manufactured exactly according to your specification
  2. your sequence, optimized with our technologies for optimal expression
  3. a sequence designed by us, based on the protein you like to express.

Our fully enzymatic process is able to use several types of DNA as input, including:

  1. circular DNA plasmid provided by you or your DNA supplier
  2. circular DNA plasmid cloned by RIBOPRO
  3. linear DNA from an de novo DNA synthesis, and/or PCR method

We synthesize the mRNA with a predefined percentage of  click-chemistry-enabled nucleotides, allowing the incorporation of a wide variety of dyes evenly-distributed over the length of the mRNA. Importantly, fluorescent-labelling of the mRNA will reduce the translational activity proportionally with the labelling degree.

Choices

When ordering custom mRNA you have various choices and options to customize the product to your exact needs. Is your favorite option not listed? Please contact us and we are happy to discuss a custom project with you.

Choice Description
Label AF647 (ex 647nm / em 670nm)
AF568 (ex 568nm / em 603nm)
AF488 (ex 488nm / em 530nm)
Other dyes available upon request
Capping Co-transcriptional (ARCA)
Enzymatic (VCE/FCE)
Capping efficiency 80-95% (ARCA)
95-99% (VCE/FCE)
Cap-type Cap-0 (for standard cell-types)
Cap-1 (for immune cells & in vivo applications)
Poly(A)tail length 0-300nt
Segmented
Amount 100 µg – 100 g
Buffer PBS (pH 7.4)
Citrate (pH 5.5)
Other buffer

How to get a quote?

Tells us via our Quotation form exactly what you want and you will receive an initial quotation as soon as possible. In the form you can also add questions, remarks and ask our RiboProfessionals™ to contact you to help make adjustments to your mRNA-design/mRNA-project. Filling out the form is fast, easy and prepares our people to support you the best they can!

Get a quote Schedule a meeting

What will happen next?

After your quotation/order request

Once you send the form in, we take the following steps a.s.a.p. to help you get the best mRNA as fast as possible:
1. a RiboProfessional™ checks your request,
2. gets in touch if needed, and
3. sends you a quotation within 24h
4. when you OK the quotation, we get started.

For repeat orders (an option in our ordering menu), you will still get a quotation, but we often already start with synthesis ‘at risk’ from DNA that is ‘on file’ to realize the shortest possible turn-around time. Of course, you may still ask for amendments of the order based on the quotation. The risk is entirely ours.

After your order

During production, we do the following things:

  1. Optimize the sequence if desired
  2. Communicate optimized sequences back to you (optional)
  3. Order DNA if needed
  4. Perform RNA synthesis
  5. Perform Quality Control
  6. Arrange shipping
  7. Check if material has arrived complete and correct
  8. Send invoice
  9. Follow-up to help you with your experiments if needed

For planning purposes, you will receive updates at steps 2, 4 and 6.

FAQ

Answers to some of your burning questions:

  • Why not synthesize mRNA yourself?

    Of course as a life-science researcher you have the knowledge and tools to produce any bio-molecule yourself. However, several issues make this difficult for (m)RNA.

    The disadvantages of self-synthesis of (m)RNA

    So also for the synthesis of mRNA; the multi-step enzyme-based reaction, with 1 or more purifications, in addition to acquiring a suitable DNA template, can be time-consuming; taking 2-3 days per synthesis.

    In addition, especially when a limited amount or number of mRNAs is required, the up-front cost for the reaction ingredients, the quality control, the certified RNAse-free buffers and consumables is high.

    But perhaps more than the reasons listed above, very few labs are well-equipped to work with such a delicate molecule, which is subject to degradation to omni-present and hard-to-destroy RNAses. Working with mRNA as a final product in an experimental setting is certainly doable, but multi-step manipulations during synthesis in a sub-optimal environment present a significant risk to the integrity of the final product.

    And finally, designing a high-performance mRNA requires very specific knowledge on 5’UTR and 3’UTR functioning, secondary structure formation of RNA, the induction of innate immunity by RNA, codon optimization, ideal A-tail length, etc.

    Why choose RiboPro custom mRNA synthesis instead?

    By ordering your desired mRNA from RiboPro’s custom mRNA synthesis service you can get started with your experiment as quickly as possible, without hassle! No need to worry about RNAse contamination during production, no need to setup complex quality control, no need to invest in the equipment and materials; we take care of it all. Our RNA experts make sure you get the best results.

  • What is included in the service?

    RiboPro’s custom mRNA service is very flexible; we include as much or as little as you need.

    Generally speaking, we have 2 flavors:
    A. You provide us with a (semi)suitable template, which we use as input for the RNA synthesis. Minor changes to the template to make it suitable are no problem for us.
    B. You provide us with the desired sequence, and we perform de novo DNA synthesis to obtain a suitable template for the RNA synthesis.

    What is included?

    So, in the full-service variant (B) the following is included:
    1. mRNA design and optional sequence optimization
    2. de novo DNA synthesis
    3. DNA prep
    4. RNA synthesis
    5. RNA capping (up to 99% capped, and Cap1 modification possible)
    6. A-tailing to desired length (up to 500nt)
    7. 2 purification round
    8. (optional) dsRNA removal procedure
    9. Quality control (length, sequence verification (DNA), A-tail length, purity)
    10. (optional) Aliquotation
    11. Packaging & shipping
    12. After-care to ensure success in your experiments

    This very complete package is available from €660 + shipment cost (typ. €50-€150) + DNA synthesis (typ. €100-€200)

  • How does RIBOPRO optimize mRNA sequences?

    The performance of a messenger RNA, to be added to cells via standard transfection methods, greatly depends on the design of the sequence, the quality of the synthesis, chemical modifications of specific bases and the purity of the end-product. Introducing a sub-optimal mRNA into cells is known to trigger the innate immune response; resulting in lower protein expression, cellular toxicity and the release of pro-inflammatory cytokines.

    High performance mRNA
    For the best performance, we recommend the following combination of features to be selected for your custom mRNA:

    • 5’UTR: no upstream ATG, kozak or similar present directly upstream of the start codon. For example RiboPro catalogue 5’UTRs.
    • 3’UTR: selected from mRNAs known to be extra-ordinary stable.
    • Cap1 – 95%.
    • 120-150nt A-tail.
    • dsRNA reduction.

    These parameters will provide you with a decent expressing mRNA. However, for any of the following situations:
    • therapeutic applications,
    • a demand for high protein-expression,
    • experiments incompatible with pro-inflammatory cytokine release,
    • or any other application requiring the best protein expression and the lowest toxicity,
    we recommend additional mRNA-specific sequence-optimization. Using RiboPro’s patent-pending technology allows for 5x more protein (as high as 40x more in specific applications) and significant reduction in pro-inflammatory cytokine production.

    If you like to understand why these settings lead to the best performance, read more below.

    The trouble avoided by RiboPro’s high-quality RNA synthesis
    Synthesizing a high-performance messenger RNA requires optimization all through-out the multi-step synthesis, including specialized purification.

    dsRNA
    First of all, during the enzymatic production process, dsRNA can be formed. dsRNA is one of the main triggers of innate immunity, primarily via TLR3 (endosomal recognition during transfection), OAS1, PKR and MDA5 (cytoplasmic sensors). RiboPro’s mRNA synthesis process results in the lowest amount of dsRNA possible. The method used to reduce the amount of dsRNA, also negatively affects the yield of ssRNA, so limited additional costs do apply compared to the standard method, because we increase the scale in order to obtain the guaranteed yield. Without dsRNA reduction, protein expression is typically 10-fold lower than with dsRNA reduction.

    Uncapped, triphosphorylated RNA
    Secondly, currently no supplier can promise that 100% of all mRNA molecules will be capped. When uncapped, the 5’UTR of the mRNA contains a 5′ triphosphate, which can be a ligand for the cytoplasmic innate immune sensor RIG-I. RiboPro offers 2 capping grades (~80% and 95%+). If selecting the lower grade (cheaper), additional phosphatase treatment may be required to reduce RIG-I activation.

    Cap methylation status
    Third, if a cap is present, but no additional methylation of the first nucleotide (Cap0), then IFIT1 may be activated in certain cells, leading to around 30-50% reduction in activity in some experiments. This seems to be particularly relevant in cells of the immune system. RiboPro offers optional methylation of the first nucleotide (Cap1).

    Contaminants
    Finally, template DNA, salts and small RNA species (formed during abortive cycling of the RNA polymerase) must be carefully removed to avoid interfering with uptake and interpretation of results. RiboPro uses extensive DNAse I-based DNA removal, reducing the residual DNA to far less than 1 millionth of the RNA concentration.

    QC
    Next to an optimized production process, RiboPro also has also developed a series of test for quality control, offering you peace of mind about the identity, purity and modification status of your order.

    Do you have additional questions? Please contact us directly below or later via the contact page. We are happy to help!

  • How fast can we deliver?

    For a typical sequence, you will receive your order within 4-5 weeks. If you provide us with a suitable template, it may even be faster, because 3rd party de novo DNA synthesis is the most time-consuming step of our RNA synthesis process.

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Custom mRNA
DNA Template*
Sequence optimization*
Purpose*

Labelled*

Per default capping is based on ARCA Active. Other capping systems (like Vaccinia) are available on request. Cap 1 is advised, unless application type is immortalized cells, then Cap 0 is advised.
5 'UTR*

3 'UTR*

Poly A tail*

Solvent*
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Nick Broos

Product Specialist

Welcome on our new website

Please bear with us while we get everything in order.
Importantly, we are ready to receive your order, quote request or meeting request as per usual.
We look forward helping you with the highest quality mRNA.