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A Simple Method to Evaluate the Length of Poly(A) Tails in mRNA

Jonas Mumenthaler1,2, Maximilian Feldmann1,2, Shahab Mamaghani1,3

  • 1Department of Dermatology, University Hospital Zürich (USZ), University of Zürich (UZH), 8091 Zürich, Switzerland.

Methods and Protocols
|July 24, 2026
PubMed

Insights

A new fluorescence-based method quickly measures the poly(A) tail length of synthetic messenger RNA (mRNA). This rapid assay simplifies quality control for mRNA vaccines and therapeutics, improving production efficiency.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmaceutical Sciences

Background:

  • Synthetic messenger RNA (mRNA) is a key component in vaccines and therapeutics.
  • The 3' poly(A) tail is crucial for mRNA stability and translation.
  • Current methods for measuring poly(A) tail length are complex and time-consuming.

Purpose of the Study:

  • To develop a faster, simpler method for determining the average poly(A) tail length in synthetic mRNA.
  • To facilitate quality control in the production of mRNA-based pharmaceuticals.

Main Methods:

  • Utilized thiazole orange, a fluorescent dye that binds to nucleic acids.
  • Developed an oligo(dT)12 oligonucleotide with thiazole orange at both ends.
  • Established a titration curve using known poly(A) RNA amounts for calibration.

Main Results:

  • The thiazole orange oligo(dT)12 probe emits fluorescence proportional to the available poly(A) sequence.
  • The method provides an instant indication of the average poly(A) tail length.
  • Demonstrated the method's applicability for in vitro-transcribed mRNA.

Conclusions:

  • This accelerated method offers a fast and easy way to evaluate synthetic mRNA poly(A) tail length.
  • The technique is suitable for any laboratory setting, supporting research and clinical applications.
  • Simplifies quality assessment for mRNA production, from research to clinical trials.
Keywords:
ECHO probein vitro transcriptionivt mRNAmRNApoly(A) tail

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