**mRNA In Vitro Transcription Service**

Quick Turnaround mRNA IVT Services for Research Grade mRNA

Molecular Cloning & Mutagenesis Projects Completed

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Viral Vector Packaging Solutions Delivered

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Plasmid DNA Preparations Successfully Produced

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Let's Begin Your mRNA Production

Our team offers mRNA IVT services on DNA sequences, up to 18 kilobase pairs including promoters, to produce and deliver high quality mRNA vectors for researchers. Although mRNA and DNA require different expertise, our production teams are capable of producing the right vector and construct to match your research applications. We offer several different RNA polymerase options including T7, SP6, and T3 to ensure your DNA template gets transcribed into mRNA correctly.

Our in vitro mRNA transcription service lead times can change, so be sure to check our current lead times.

Getting Started on mRNA IVT Transcription

mRNA transcription services can start in several different ways:

Option 1: Open Reading Frame (ORF): Researchers can ship their ORF clones to our facility for molecular cloning into a backbone and promoter insertion such as: T7, SP6, or T3. Note: Our facility does not offer gene synthesis however we are able to refer researchers to certain vendors.
Option 2: Plasmid DNA: Shipping a fully designed plasmid (with or without additional molecular cloning or sequence mutation updates) is the simplest, most cost effective, and decreases production time for researchers. Additionally, plasmid DNA is highly stable and ships well.
Option 3: RNA construct: Although mRNA cannot be used as a template for IVT, our team is able to use RT-PCR to resynthesize a complicated template. This technique is time and resource consuming compared to the above options.
Option 4: PCR template: Ideal for high volume quick small batch productions of mRNA, researchers can send a PCR product with promoter and poly(A) included for high volume quick turnaround mRNA production.

mRNA In Vitro Transcription Workflow

Template Design and Preparation (If using Options 3 or 4, skip to the next section): Depending on which starting material is used and delivered to our production facility, our team starts by creating a pDNA template using an ORF or whole construct. The plasmid carrying a GOI carrying a T7, SP6, or T3 promoter is grown in E. coli, purified, and then linearized with a restriction enzyme right after the poly(A) tail.
IVT reaction: The linear DNA template is combined with RNA polymerase, NTPs, a cap analog if co-transcriptional capping is used, magnesium, and buffer. The polymerase binds the promoter and synthesizes RNA along the template. Our team has extensive experience working with linearized, circular, highly (positive or negative) supercoiled plasmid DNA.
Purification and QC: DNase treatment removes the template, and the mRNA is then purified, validated using analytical QC assays including gel electrophoresis, RT-qPCR, and Sanger sequencing. The final mRNA can be packaged into LNPs or other particles depending on the researcher’s application(s).

Production Options

Our mRNA in vitro transcription services offer 3 different polymerase options to choose from. T7 remains the most common promoter, however, if you are unsure which promoter is best for your research application contact our team.

Choosing the promoter early is highly recommended because it affects downstream template design. The promoter in the template and the enzyme in the reaction must match. The template doesn’t hold the enzyme, it only holds the promoter.

Here is a quick breakdown of the three promoters we offer:

Typical role	Default for research and therapeutic mRNA	Alternative to T7	Rarely used for mRNA
	T7	SP6	T3
Promoter (core)	TAATACGACTCACTATAG	ATTTAGGTGACACTATAG	AATTAACCCTCACTAAAG
Yield	Highest	Moderate	Moderate to lower
Commercial support	Extensive, including engineered variants and capping reagents	Good	Limited

Additional Notes:

T7 gives the best yields and is the best characterized
SP6 and T3 are useful when a template needs to avoid T7 activity. A common example is dual-promoter plasmids that make sense and antisense RNA from opposite ends.
Each enzyme has its own termination and pausing tendencies, so a template that terminates prematurely with T7 may behave better with SP6.

Sample Submission Requirements

Depending on how you wish to start your mRNA transcription production, please select from one of the following options:

Open Reading Frame (ORF)
- Requires 3 µg and full sequence of the template
Plasmid DNA
- Requires 3 µg, full sequence of the template, plasmid map, and areas of antibiotic resistance
- Any additional cloning or mutation requests
RNA construct:
- Requires 10 µg
PCR template
- Requires 5 µg

mRNA Vector Deliverable

5 ug of mRNA
Electronic project report with DNA-to-mRNA sequence alignments and QC assay results

mRNA Integrated Vector Sequencing Reports for IND Filing

Integrated Vector Sequencing Reports are a critical component of any Investigational New Drug application submitted to the FDA for gene therapy products. This analysis provides essential verification of the vector’s genetic integrity, identity, and stability. By documenting the complete nucleotide sequence of a therapeutic vector, identifying any potential sequence variants, and confirming genetic stability across manufacturing processes, this report addresses crucial regulatory requirements while demonstrating a commitment to product quality.

Our mRNA team can provide Integrated Vector Sequencing Reports for mRNA vectors, just ask when starting a project.

mRNA in Research and Development

Want to learn more about the latest in mRNA? Our colleagues at ScienceDirect, the American Society for Biochemistry and Molecular Biology, and Genetic Engineering & Biotechnology News continuously collect and publish the latest information on mRNA.

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mRNA In Vitro Transcription Service