mRNA in vitro transcription

 Service Introduction

        mRNA therapeutics have been recognized as a promising gene therapy platform with unlimited potential to revolutionize modern medicine. Compared to recombinant protein production, mRNA production through in vitro transcription is faster, more flexible, and less expensive. In the past decade, significant advances in mRNA chemical modification and delivery systems have rapidly accelerated the application of this technology in clinical research. mRNA has a wide range of potential applications. In addition to being used to prevent infectious diseases, mRNA vaccines also bring new hope for the treatment of cancer and immune diseases.

        IGEBio has developed a plasmid that can carry a long polyA chain of 100-150 bp and can be used for in vitro transcription. The linearized plasmid is used to prepare mRNA by T7 in vitro transcription, and the gene length can be as long as 2 kb to 3 kb. After a series of specific process flows such as capping and magnetic bead purification, our in vitro-prepared mRNA molecules can be customized and optimized for different downstream experiments, such as cell transfection, embryo injection, and biological drug and vaccine development.

Service Advantages

        Use T7 RNA polymerase derived from bacteriophages to efficiently synthesize mRNA molecules with a length of more than 3 kb.

        Add a 5' m7G cap and a 3' polyA tail of 100nt to the mRNA molecule. The polyA tail can be extended to 100 to enhance the stability and translation efficiency of mRNA.

        Provide free codon optimization for different species to enhance mRNA translation efficiency.

        The transcription product is treated with DNase and phosphatase to remove residual DNA template and 5' phosphate group of mRNA.

Vector map

Synthesis process and verification

Experimental verification

        Our in vitro-transcribed mRNA has been specially optimized to enhance stability and help protein translate efficiently. The following figures show the cases of successful in vitro transfection of HEK293T cells with EGFP mRNA designed by us and the electrophoresis of purified mRNA:

293Ta-mRNA-0.5ug-transfect-48h-100x-1

Cap 1-capped EGFP mRNA transfection HEK293T cells-1

Cap 1-capped EGFP mRNA transfection HEK293T cells-2

In vitro transcription principle

Left figure: Relationship between reaction time and yield

Right figure: Relationship between template input and yield

Transportation/storage 

        Dry ice transportation, store the sample at -80℃ after receiving it.

Notes

        IGEBio's mRNA is stored in enzyme-free water and transported in dry ice form.

        After receiving the goods, please place the mRNA at -80℃ for long-term storage (can be stably stored for at least 6 months), or place it at -20℃ for one week.

        The shelf life of mRNA is 1 year. Do not freeze and thaw mRNA repeatedly, otherwise it will degrade significantly.