3AIM-seq: Quality assessment of mRNA therapeutics using sequencing for 3' poly(A) tails of in vitro-transcribed mRNA.

ABSTRACT: In vitro-transcribed (IVT) mRNA therapeutics are promising for preventing and treating diseases, including infectious diseases and cancer, by delivering nucleic acid sequences. Assessing the stability of mRNA sequences and poly(A) tail length is crucial to minimize adverse effects and ensure drug efficacy. However, accurately measuring long homopolymeric nucleotides remains technically challenging, and a specialized method for IVT mRNAs is lacking. We introduce 3AIM-seq, a technique optimized experimentally and analytically for sequencing 3' poly(A) tails in IVT mRNAs. To generate high-quality data, we used a ligation-free adapter followed by 3' end amplification to prepare high-throughput sequencing libraries. The 3AIM-seq algorithm employed a sliding window approach to base quality score

SUBMITTER: Seo J 

PROVIDER: S-EPMC12848163 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature