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As a trendy genetic medicine, in vitro transcribed mRNA (IVT mRNA) has several advantages for gene delivery including no risk of insertional mutagenesis, streamlined cell-free manufacturing, and the ability to develop personalized treatments. IVT mRNA can be applied to a wide range of therapeutic applications including vaccines, protein replacement, CAR-T, and CRISPR gene editing. VectorBuilder's expert RNA team specializes in custom IVT mRNA design, production, and LNP encapsulation, with a wide range of customization possibilities.

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Royalty-free IVT backbone for mRNA production with no IP constraints for commercial use


As fast as 5 weeks from vector cloning
to LNP encapsulation


Comprehensive customization including capping methods, modified nucleotides, poly(A) tails, and UTRs


Expert design and production team specialized in optimizing your mRNA
for the highest expression, yields, and quality

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Technical Information

The following diagram depicts the fundamental components of mRNA, and each component plays critical roles in regulating gene expression. There are a variety of methods for assembling these components in vitro and VectorBuilder can help assess the best method for achieving optimal expression, yields, and purity for your project. You can also read the Vector Academy article The Basics of mRNA Therapeutics for a more detailed overview of therapeutic mRNA.


Figure 1. Structure and function of mRNA components.


How do mRNA, circRNA, and saRNA compare to each other?
mRNA circRNA saRNA
Structure Linear; usually contains 5’ cap, 5’ UTR, ORF for GOI, 3’ UTR, and poly(A) tail Circular; usually contains IRES and ORF for GOI Linear; usually contains 5’ UTR, ORFs for replicase genes and GOI, 3’ UTR, and poly(A) tail
Cap Requires 5' Cap for stability and ribosome recruitment No Cap; relies on IRES for ribosome recruitment Requires 5’ Cap for stability and ribosome recruitment
RNA length (nt) 100 ~ 10,000 1000 ~ 5000 7000 ~ 10,000
Stability Low High Low
Can modified nucleotides be added in production? Yes No Yes
Expression level Low Medium High
Expression duration Short Medium Long
Immunogenicity Low Medium High
What are the differences between mRNA caps and capping methods?

Cap 0 refers to N7-methylguanosine (m7G) that is added to the 5’ end of eukaryotic mRNAs via a 5’ to 5’ triphosphate linkage. This modification is added via a series of enzymatic reactions that occur co-transcriptionally and functions to regulate nuclear export, transcript stability, and promotes translation of the mRNA through recognition by eukaryotic translation initiation factor (eIF4E). Cap 1 refers to the addition of a methyl group to the 2’O on the first nucleotide (m7GpppNm) of the transcribed mRNA sequence in addition to the m7G cap. In mammalian cells, cap 1 structure is an important marker for mRNA to be recognized as self and not targeted by innate immunity. Adding cap 1 structure to synthesized mRNA has been demonstrated to enhance mRNA expression in vivo and reduced its immunogenicity.

Capping for in vitro transcribed RNA can occur either co-transcriptionally with cap analogs or post-transcriptionally via enzymatic reactions. We offer both capping methods, and the efficiency of them has been well validated using LC-MS. Depending on the client-preferred capping method, we will choose a compatible backbone for cloning the IVT mRNA vector from the beginning.

Why should I consider incorporating modified nucleotides in mRNA and which ones can be included?

Cells contain cytosolic and endosomal RNA receptors that activate the immune response upon recognition of foreign RNA. Modified nucleotides are commonly found in endogenous cellular RNA. Incorporating certain modified nucleotides in IVT mRNA reduces its immunogenicity, alters secondary structure, and increases translation efficiency and half-life in a sequence-dependent manner. We provide a wide range of modified nucleotides, including the commonly used N1-Methylpseudouridine (m1Ψ) and 5-Methylcytosine (m5C). N1-Methylpsuedouridine and 5-Methylcytosine are naturally occurring nucleotides that were first identified in tRNAs, however, their use in coding mRNAs has only recently been appreciated. These methylated derivatives of uridine and cytosine can replace their canonical nucleotides in mRNA IVT and translation without altering traditional Watson-Crick base pairing. A major advantage to their use in mRNA therapeutics is their ability to alter recognition by RNA immune receptors thus mitigating unwanted immune effects and enhancing transcript stability and translation.