Accelerate Your mRNA Research

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

Step 1Gene synthesis and plasmid production
Step 2mRNA synthesis
Step 3mRNA purification and delivery
Step 4mRNA effect monitoring
Step 5Measure protein expression

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.

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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.

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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.

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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.

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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.