In vitro transcription with a T7 RNA polymerase.
Messenger RNA (mRNA) synthesis by in vitro transcription (IVT) is a cell-free process to produce RNA by enzymatic reactions. Synthesized RNA such as mRNA is widely utilized in research, drug development, vaccine production, and many other applications.
Key components of in vitro transcription are:
- Linear DNA as a template
- An RNA polymerase as a synthesizing enzyme, transcribing RNA from DNA
- Nucleotides as building blocks, which may be modified or unmodified
- Modifying enzymes for 5′ capping and 3′ poly-A tailing (for eukaryotic mRNA)
An RNase inhibitor and other components like pyrophosphatase may be added to the IVT reaction to help improve RNA yield.
Capping and tailing of mRNA.
Table 1. Key products for mRNA synthesis by in vitro transcription.
| DNA template generation | RNA synthesis | Post-transcriptional modification | DNA template and protein digestion |
|---|---|---|---|
Scaling mRNA synthesis for different applications
The IVT reaction can be scaled to produce mRNA from micrograms to milligram, or even gram scale and beyond, for the amount needed for your intended application. For manufacturing or production of RNA at large scales for therapeutic development, quality standards and regulatory requirements of the raw materials must be considered.
Thermo Fisher Scientific offers a comprehensive portfolio of in vitro transcription products to meet the requirements for the entire spectrum of mRNA applications—from research to therapeutic development and vaccine production. Many of these products are available individually or as kits designed to facilitate in vitro transcription.
Table 2. Recommended IVT solutions by mRNA synthesis scale.
| Scale | Micro scale | Mini scale | Mid-scale | Production scale |
|---|---|---|---|---|
| Volume | μLs to 200 mL | Up to 500 mL | 1–5 L | 10–50 L |
| mRNA yields (from multiple runs per year) | Microgram to milligram range | Milligram range | Milligram to gram range | Gram to kilogram range |
| Applications |
|
|
| Commercial manufacture |
| Performance needs | High-throughput | Reproducible | Scalable | Reliable |
| Product quality support | For Research Use Only | For Research Use or Further Manufacturing | For Research Use or Further Manufacturing | For Research Use or Further Manufacturing |
| Recommend solutions |
Featured products for mRNA research
MEGAscript T7 Transcription Kit Plus
Next-generation in vitro transcription kit for uncapped mRNA synthesis
mMessage mMachine T7 mRNA Kit with CleanCap Reagent AG
In vitro transcription kit designed with third generation capping technology for superior capped mRNA synthesis
Featured products for mRNA production
Thermo Scientific TheraPure GMP* enzymes and nucleotides
Designed to meet critical process, scale, quality, and regulatory requirements for large-scale mRNA production of therapeutics and vaccines
Invitrogen Dynabeads mRNA production solutions
Designed to support scalable GMP-grade synthesis and purification of mRNA for vaccine development and manufacture
Resources
mRNA production
* “TheraPure GMP” refers to the quality level of the raw, ancillary, or starting materials to be used for further manufacturing. TheraPure GMP products are manufactured in facilities with ISO 9001–certified quality management systems that operate in accordance with relevant good manufacturing practice (GMP) principles, as outlined in ICH Q7 or equivalent guidance documents or standards.
Intended use of the products mentioned on this page vary. For specific intended use statements please refer to the product label.