In vitro transcription basics
Dive into in vitro transcription with an experimental overview and resources to understand how mRNA is synthesized.
A proven end-to-end product portfolio for mRNA research
The process for mRNA research and discovery requires flexible products and technologies to meet many of your diverse needs. To effectively facilitate mRNA discovery, Thermo Fisher Scientific offers a variety of products that span the entire portfolio from target gene identification and synthesis, plasmid production, in vitro transcription, and delivery through tomeasuring protein expression and immune response.
mRNA discovery workflow
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Target genes for mRNA therapeutics and vaccines are often discovered and validated using techniques, such as next generation sequencing (NGS). The target sequence will be selected, optimized, synthesized, and inserted into a DNA plasmid to use as a template for mRNA synthesis using in vitro transcription to produce the mRNA for further testing.
Featured products
- Ion Torrent Next Generation Sequencing Systems
- Invitrogen GeneArt Gene Synthesis
- Thermo Scientific PCR enzymes and master mixes
Resources
In vitro transcription uses the linearized DNA template containing the target gene, nucleotides, and enzymes to synthesize mRNA. The resulting mRNA product can be synthesized utilizing modified nucleotides or with mRNA capping and tailing to enhance protein translation, improve stability, or reduce immunogenicity as required.
Featured products & services
In vitro transcription
- Invitrogen mMessage mMachine T7 mRNA Kit with CleanCap® Reagent AG
- Invitrogen mMessage mMachine T7 Ultra Transcription Kit
- MEGAscript T7 Transcription Kit Plus
- MEGAscript T7 Transcription Kit
- Custom mRNA Synthesis Services
- Dynabeads Streptavidin for In Vitro Transcription
mRNA clean-up
Quantification
Resources
The final mRNA product is isolated and purified by affinity purification using beads or a column and further prepared using chromatography and filtration as needed. After purification, the mRNA is formulated for delivery to the test model system typically using lipid nanoparticles that are made up of a number of component lipids and other materials.
Featured products
Purification
- Dynabeads Carboxylic Acid for RNA Purification
- Dynabeads mRNA Purification Kits
- Kingfisher Sample Purification Systems
- MEGAclear-96 Transcription Clean-Up Kit
Delivery
- Invitrogen Vivofectamine Delivery Solutions
- Lipofectamine MessengerMAX mRNA Transfection Reagent
- Transfection Reagents
Quantification
- NanoDrop Eight Spectrophotometer
- Qubit Fluorometric Quantification
- E-Gel Precast Agarose Electrophoresis System
Resources
Once the target mRNA has been synthesized, formulated, and delivered, measuring uptake and incorporation of the mRNA is typically done using a cell- or tissue-based system. mRNA incorporation can be measured and visualized using several techniques including RT-PCR, Northern blot, or in situ hybridization.
Featured products
- Total RNA extraction kits
- RT-PCR reagents and kits
- Northern blot reagents and systems
- In situ hybridization
Resources
mRNA therapeutics or vaccines are often designed to induce production of a protein or protein fragment in target cells and patients. Measuring the production of the protein encoded by the mRNA is key to determining the success of a mRNA therapeutic candidate. During the pre-clinical phase, protein expression can be measured indirectly using techniques such as western blotting or fluorescence detection, or directly by liquid chromatography-mass spectrometry (LCMS).
T cell activation is also an important measure of the activity of an mRNA, particularly with immunotherapies, and antigen processing and presentation can be monitored using appropriate T cell gene expression microarrays.
Alternatively, many mRNA based immune therapies are designed to stimulate humoral immunity through the production of antibodies directed at the target protein encoded by the mRNA. Measuring this antibody response to the mRNA therapeutic candidate is vital information for an mRNA immunotherapeutic or vaccine. During the discovery phase, ELISA assays are a commonly used technique to track an antibody or protein response to an mRNA therapeutic candidate.
Featured products
Kits and assays
Imaging systems
- Thermo Scientific Varioskan LUX Microplate Reader
- Applied Biosystems TaqMan Array Human Antigen Processing and Presentation by MHCS
- Invitrogen iBright FL1500 Imaging System
- Attune Flow Cytometers
Resources
Target genes for mRNA therapeutics and vaccines are often discovered and validated using techniques, such as next generation sequencing (NGS). The target sequence will be selected, optimized, synthesized, and inserted into a DNA plasmid to use as a template for mRNA synthesis using in vitro transcription to produce the mRNA for further testing.
Featured products
- Ion Torrent Next Generation Sequencing Systems
- Invitrogen GeneArt Gene Synthesis
- Thermo Scientific PCR enzymes and master mixes
Resources
In vitro transcription uses the linearized DNA template containing the target gene, nucleotides, and enzymes to synthesize mRNA. The resulting mRNA product can be synthesized utilizing modified nucleotides or with mRNA capping and tailing to enhance protein translation, improve stability, or reduce immunogenicity as required.
Featured products & services
In vitro transcription
- Invitrogen mMessage mMachine T7 mRNA Kit with CleanCap® Reagent AG
- Invitrogen mMessage mMachine T7 Ultra Transcription Kit
- MEGAscript T7 Transcription Kit Plus
- MEGAscript T7 Transcription Kit
- Custom mRNA Synthesis Services
- Dynabeads Streptavidin for In Vitro Transcription
mRNA clean-up
Quantification
Resources
The final mRNA product is isolated and purified by affinity purification using beads or a column and further prepared using chromatography and filtration as needed. After purification, the mRNA is formulated for delivery to the test model system typically using lipid nanoparticles that are made up of a number of component lipids and other materials.
Featured products
Purification
- Dynabeads Carboxylic Acid for RNA Purification
- Dynabeads mRNA Purification Kits
- Kingfisher Sample Purification Systems
- MEGAclear-96 Transcription Clean-Up Kit
Delivery
- Invitrogen Vivofectamine Delivery Solutions
- Lipofectamine MessengerMAX mRNA Transfection Reagent
- Transfection Reagents
Quantification
- NanoDrop Eight Spectrophotometer
- Qubit Fluorometric Quantification
- E-Gel Precast Agarose Electrophoresis System
Resources
Once the target mRNA has been synthesized, formulated, and delivered, measuring uptake and incorporation of the mRNA is typically done using a cell- or tissue-based system. mRNA incorporation can be measured and visualized using several techniques including RT-PCR, Northern blot, or in situ hybridization.
Featured products
- Total RNA extraction kits
- RT-PCR reagents and kits
- Northern blot reagents and systems
- In situ hybridization
Resources
mRNA therapeutics or vaccines are often designed to induce production of a protein or protein fragment in target cells and patients. Measuring the production of the protein encoded by the mRNA is key to determining the success of a mRNA therapeutic candidate. During the pre-clinical phase, protein expression can be measured indirectly using techniques such as western blotting or fluorescence detection, or directly by liquid chromatography-mass spectrometry (LCMS).
T cell activation is also an important measure of the activity of an mRNA, particularly with immunotherapies, and antigen processing and presentation can be monitored using appropriate T cell gene expression microarrays.
Alternatively, many mRNA based immune therapies are designed to stimulate humoral immunity through the production of antibodies directed at the target protein encoded by the mRNA. Measuring this antibody response to the mRNA therapeutic candidate is vital information for an mRNA immunotherapeutic or vaccine. During the discovery phase, ELISA assays are a commonly used technique to track an antibody or protein response to an mRNA therapeutic candidate.
Featured products
Kits and assays
Imaging systems
- Thermo Scientific Varioskan LUX Microplate Reader
- Applied Biosystems TaqMan Array Human Antigen Processing and Presentation by MHCS
- Invitrogen iBright FL1500 Imaging System
- Attune Flow Cytometers
Resources
Custom mRNA Synthesis Service
Our Custom Biology Services group is ready to help build the mRNA to meet any of your small-scale research needs. Whether you have a full mRNA template ready to go or want to plug-n-play with our proven designs, our flexibility in scales and QC options can support a variety of applications. We offer fully customizable mRNA with 5’ cap options, including ARCA or CleanCap® Reagent AG, modified nucleotides, and scales as low as 50 μg. We support the entire workflow from in vitro transcription template design through mRNA synthesis and lipid nanoparticle encapsulation, with a seamless transition into our cGMP manufacturing offerings.
To submit an inquiry for our Custom mRNA Synthesis Service, use our general service inquiry form. Simply click below to navigate to the form, and check the box for Custom mRNA Synthesis Services within the Cell Engineering tab.
Learn more about in vitro transcription
History of in vitro transcription
Discover the evolution of mRNA research and in vitro transcription, from foundational discoveries through modern-day implications.
Discover mRNA research applications
mRNA applications in oncology
Understand how synthetic mRNA produced via in vitro transcription has emerged as a useful tool in oncology therapeutic research.
In vivo applications
Learn how mRNA vaccines are utilized in vivo and the role of lipid nanoparticles.
mRNA vaccine developments
Uncover how COVID-19 vaccine development has influenced mRNA vaccine research in other therapeutic areas.
Support
For Research Use Only. Not for use in diagnostic procedures.