In vitro transcription (IVT) mRNA is widely used in basic research and application which provides a promising gene expression system for rapid and controllable induction of target proteins. Based on its safety and high efficiency, IVT mRNA is a powerful approach with potential applications in regenerative medicine, vaccination, cell engineering, and protein replacement therapy. IVT mRNA synthesis with internal epigenetic modifications requires a DNA template, enzymes, nucleotides, and buffer components. Not familiar with IVT mRNA technology or looking for a fast and effective service? Abnova provides full mRNA synthesis service encompassing DNA template production, mRNA production, and mRNA QC to fulfill customer demand. Our technical support is available to accommodate additional requests and needs required by your experiments.

mRNA Synthesis Flowchart

 

Advantages

  • Customized mRNA from several hundred to thousands of bases
  • Optimized procedure of mRNA synthesis and purification
  • Cell-free production without toxic chemicals
  • Qualified RNA to match customer needs
  • Ready for most downstream applications
  • High-quality services with competitive prices
 
 

Specification

IVT Synthesis of mRNA Service Details


Services Items

Lead Time

DNA Template Production

Amplification of the gene of interest* and cloning it into the expression vector

4 weeks

Plasmid DNA preparation and purification

mRNA Production

Linearized DNA preparation

4 Days

In vitro transcription of linearized DNA into RNA

RNA capping

Capped RNA purification and quantification**

mRNA QC

Integrity and purity: agarose gel electrophoresis of RNA

2 Days
Protein expression: RNA transfection into cell and cell lysate subjected to Western blot analysis*** 2 Weeks
 

          Note:

          * Customers provide target sequence or DNA plasmid
          ** Adjustable amount of RNA yield to match customer demand
          *** Optional task depends on customer needs

 

 

Capping Methods

 

Capping methods

Co-transcriptional Modification
Sequence

RNA Sequence

Capping sequence Cap Analog Features
Cap 0

GAUG

3´-O-Me-m7G(5')ppp(5')GAUG Anti-Reverse
GTP competition
Cap 1

AUG

m7G(5')ppp(5')(2'OMeA)pUG Without Anti-Reverse
Without GTP competition
Cap 1 AGG m7(3'OMeG)(5')ppp(5')(2'OMeA)pGG Anti-Reverse
Without GTP competition
Cap 1 GGG m7(3'OMeG)(5')ppp(5')(2'OMeG)pGG Anti-Reverse
GTP competition
Post-transcriptional Modification
 Sequence  RNA Sequence Capping sequence 
Cap 0 AUG
UUG
GUG
CUG
m7G(5')ppp(5')AUG
m7G(5')ppp(5')UUG
m7G(5')ppp(5')GUG
m7G(5')ppp(5')CUG
Cap 1 AUG
UUG
GUG
CUG
m7G(5')ppp(5')(2'OMeA)UG
m7G(5')ppp(5')(2'OMeU)UG
m7G(5')ppp(5')(2'OMeG)UG
m7G(5')ppp(5')(2'OMeC)UG
Cap 1 AGG
UGG
GGG
CGG
m7G(5')ppp(5')(2'OMeA)GG
m7G(5')ppp(5')(2'OMeU)GG
m7G(5')ppp(5')(2'OMeG)GG
m7G(5')ppp(5')(2'OMeC)GG
Cap 1 ACG
UCG
GCG
CCG
m7G(5')ppp(5')(2'OMeA)CG
m7G(5')ppp(5')(2'OMeU)CG
m7G(5')ppp(5')(2'OMeG)CG
m7G(5')ppp(5')(2'OMeC)CG
 
 

Nucleoside Modifications

  • Pseudouridine (Ψ)
  • 5-methylcytidine (m5C)
  • N1-methylpseudouridine (N1mΨ)
 
 

Improved mRNA Expression with Cap1 and Pseudouridine Modification

IVT_mRNA_Ex1.jpg (35 KB)

 

 

Improved SAM Expression with Cap1 without Nucleoside Modification

IVT_mRNA_Ex2.jpg (39 KB)

 

 

Improved mRNA Expression with Polyadenylated PolyA

IVT_mRNA_Ex3.jpg (41 KB)

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