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

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.