The Rush Is Real

Case Studies: Faster mRNA, Proven Results.

• High Protein Expression Level

  Rush mRNA has similar protein expression levels compared to regular mRNA and can be delivered faster while ensuring downstream applications.

  
• New Cap Analogue 2′-O-Ethyl-Capped RNA Enhances Half-Life In Vivo

  Novel 2’-O-Ethyl-capped RNA showed higher expression than natural Cap1 RNA, and slower clearance rate in vivo.

  

  

FAQs

• What template does GenScript use for mRNA manufacture? How is that made?

  We use linearized DNA plasmid as a template for manufacturing your mRNA. The gene can be designed by our team using GenScript’s proprietary vectors and tools or we can use one of your designs. Once the design has been agreed to, we handle the gene and mRNA manufacture and provide you with mRNA as part of the fee-for-service agreement.

• What 5’ cap does GenScript offer?

  Cap1 via co-transcriptional capping is our default recommendation. We can also offer Cap0 and Cap1 enzymatically for an additional cost, and Cap2 RNA through co-transcriptional capping technology.

• How is the capping efficiency of GenScript's mRNA platform?

  GenScript's proprietary capping technology can achieve >98% capping efficiency.

• How should I store my mRNA?

  mRNA should be stable for at least 1 year if stored properly at -80°C upon receival and avoiding repeated freezing and thawing cycles, however mRNA stability varies depending on the length of mRNA. We have tested mRNA with stable results up to 3 freeze/thaw cycles. We recommend storing at -80°C for long time storage and -20°C for short term storage.

• What is the length limit of GenScript's mRNA production?

  We can make mRNA from 100bp to 20kb in length. For oligo shorter than 100bp, our oligo synthesis team can take it under chemical syntehsis.

• What’s the difference between RUO and preclinical grade?

  For RUO grade mRNA, we use silica membrane or LiCl precipitation method for purification. It is suitable for in vitro and invivo proof of concept studies.

  For preclinical grade, we use HPLC based Oligo dT purification for improving RNA purity. Preclinical grade can be considered for long mRNA constructs >5kb or when high RNA purity (less fragment RNA ) is required for the project.

• Can GenScript provide GMP mRNA ?

  We can provide cGMP mRNA through our sister company ProBio. Please contact our technical support team.

• Do you guarantee a specific mRNA yield/amount?

  Yes. We guarantee specific amounts of mRNA be delivered.

• What concentration can you ship my mRNA at?

  We ship mRNA at concentration of 0.5 ~2mg/mL by default. We can adjust concentration to a specific value <4mg /mL per customer's request at extra cost.

• What buffer options does GenScript provide for mRNA ?

  We offer 3 options of buffer: 1mM sodium citrate pH 6.5, TE pH7, and RNase-Free H2O. 1mM sodium citrate is our default option for better mRNA stability.

• How does GenScript purify mRNA?

  For RUO grade mRNA, we use silica membrane or LiCl precipitation method for purification.

  For preclinical grade, we use HPLC-based Oligo dT purification for improving RNA purity.

  In addition, we offer optional dephosphorylation to further reduce triphosphate RNA species in the final product to reduce its immunogenicity, and optional dsRNA removal HPLC purification to provide higher quality mRNA if low dsRNA residue is required.

• What method does GenScript use for dsRNA removal?

  Cellulose based HPLC method.

• How to define the purity for mRNA?

  For size-based purity, we run the mRNA sample on bioanalyzer, then calculate the ratio of the main peak (target size +-15%) over the total AUC.

• What’s the function of dephosphorylation?

  We recommend customers dephosphorylation as reducing the triphosphate-RNA species in mRNA by dephosphorylation could reduce its immunogenicity, thus making the transfected cells healthier and can express the target protein better.

• What’s the function of modification of mRNA?

  Modifications help stabilize mRNA and reduce its immunogenicity.

• What modified bases does GenScript offer?

  We can substitute uridine bases with either 5-moU, N1-me-Ψ, Ψ, 5’-iodourdine, or Cy5-UTP.

  We can substitute cytidine bases with 5-me-C, 5’-iodocytidine, or thiol-CTP.

  We can substitute customizable percentage substitutions upon request. These will be not site specific modifications.

• What’s the function of capping?

  Capping can stablize RNA from 5'end and recruit ribosome. Cap1 and Cap 2 have a higher impact on mRNA stability compared to Cap 0 regarding protecting the mRNA from exonuclease degradation. It is common to use Cap1 for mammalian cells and Cap 0 for plant cells. Cap2 is a novel solution provided by GenScript that may work better than Cap1 in mammailan cells especiailly in immune-stressed conditions.

• Does GenScript provide Cap Efficiency analysis as stand-alone service?

  Yes, we offer mRNA QC service including capping efficiency assay using enzyme digestion /LC-MS method. Please contact our technical support team for details.

• Can I use my own UTRs for GenScript to produce mRNA?

  Yes, it is possible, please contact our technical support team.

• What is the requirement for T7 promoter?

  For making cap1 mRNA using co-transcriptional capping technology, either AGG or GGG following T7 promoter seq is required. For self-amplifying RNA usually ATG after T7 promoter.

• Does GenScript provide mRNA codon optimization?

  Yes, we can do codon optimization. Please contact our technical support team.

• Can you use plasmids I have already made with GenScript in previous gene synthesis orders for mRNA production?

  Yes. we can subclone your ORF sequences into our proprietary vector which contains T7 promoter, UTR and 100A for mRNA production.

• For the poly A length assay, what enzyme does GenScript use?

  We use RNaseT1.

• Can GenScript perform quality testing on customer-provided mRNA?

  Yes, please refer to https://www.genscript.com/mrna-qc-services.html.

• How much mRNA is needed to transfect cell cultures in 24-well plates?

  Approximately 0.5ug to 1 ug of mRNA is usually used for transfecting each well in 24 well plate.

Resources

Questions On Storage And Usage Guidelines_mRNA, saRNA and LNP

CRISPR Knock-in Protocol for HEK 293T/293 cells with Thermo Fisher Neon® Transfection System

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