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mRNA Sequence Design

mRNA Sequence Design

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mRNA Sequence Design

According to the central dogma, messenger RNA (mRNA) is the bridge for the transmission of genetic material from DNA to proteins.

mRNA plays a biological role by encoding proteins in vivo, and mature mRNA in eukaryotic organisms consists of five components: 5' Cap (cap structure), 5' UTR (non-coding region), the ORF (open reading frame), 3ʹ UTR, and 3' polyA tail (polyadenylate tail).

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Services Details
Process Optional Service Service Details Delivery Period (Day)
mRNA sequence design and optimization Design and optimization of coding sequences

CDS sequence alignment

CDS codon optimization

 1
Design and optimization of non-coding sequences

5' UTR sequence design and optimization

3' UTR sequence design and optimization

polyA sequence design and optimization

 1-2
Customizable Options
5’ UTR/3’ UTR
  • Nature UTR sequence
  • Mutant/Engineered UTR sequence
3' PolyA Tail
  • 100A ~120A Tail (recommended)
  • Segmented polyA tail
  • Other Custom tail
Common Strategies for mRNA Sequence Design
mRNA Components Biological Functions Optimization Strategies
5’ Cap Protect mRNA from degradation by exonucleases and act in concert with the polyA tail at the 3' end, polyA binding protein and translation initiation factor protein to initiate protein translation. The natural Cap1 structure avoids pattern recognition receptors and thus reduces the natural immune response, which can be achieved by one-step co-transcriptional capping or two-step enzymatic capping [see mRNA enzymatic capping and co-transcriptional capping for details].
5’ UTR The 5' UTR can be recognized by ribosomes, regulate the translation of mRNA and affect the stability of mRNA. Contain Kozak sequences without a very stable secondary structure. Natural UTRs of highly expressed genes are preferred for in vitro transcription (IVT) mRNAs such as α-globin and β-globin.
CDS Protein-coding regions and coding sequences for antigens, antibodies, or other functional proteins. Codon optimization increases the level of translation, noting that certain non-optimal codons may play a role in protein folding.
3’ UTR Regulate mRNA translation and stability. Natural UTRs of highly expressed genes are preferred for IVT mRNAs such as α-globin and β- globin.
3’ polyA tail Regulate protein expression and protect cap structure from degradation. Adequate length (100-150 bp) is required; encoding polyA tail on the transcription template plasmid ensures a more defined polyA tail length.
Our Features
  • Diversified UTR source selection

Multiple sources of highly expressed natural & modified UTR libraries; mature UTR modification strategy;

  • Cutting-edge CDS optimization team

Cooperate with a professional AI algorithm team to complete the optimization of codons.

  • Even polyA tail distribution

Add polyA sequences according to DNA templates to control mRNA length more precisely.

  • Diversified optimization combinations

Achieve efficient expression of mRNA with low immunogenicity.

Case study

Sequence Design of a Dual-reporter mRNA: mCherry-eGFP mRNA

Yaohai Bio-Pharma’s mRNA service continues to be upgraded with the design and optimization of a double reporter gene tandem sequence, which achieves co-expression of dual genes.

Using a conventional transfection reagent, the double gene tandem sequence mCherry-eGFP mRNA is transfected into 293T cells, and two fluorescent signals of mCherry (red) and enhanced green fluorescent protein (eGFP) are detected with simultaneous expression after 48 hours, and the stacked graph is highlighted in yellow.

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Expression of mCherry-eGFP mRNA in 293T cell

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