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Sequencing of Transcriptome-Wide Poly(A) Tails by PAIso-seq
Yusheng Liu1, Falong Lu2,3, Jiaqiang Wang4
1College of Life Science, Northeast Forestry University, Harbin, China.
Methods in Molecular Biology (Clifton, N.J.)
|October 12, 2023
Summary
Poly(A) tail sequencing is now feasible with the novel PAIso-seq method. This technique accurately measures poly(A) tail length and base composition from total RNA, advancing transcriptome analysis.
Area of Science:
- Molecular Biology
- Genomics
- RNA Biology
Background:
- Poly(A) tails are crucial for eukaryotic mRNA regulation.
- Obtaining sequence information from poly(A) tails is difficult due to their homopolymeric nature.
Purpose of the Study:
- To describe poly(A) inclusive full-length RNA isoform-sequencing (PAIso-seq).
- To enable transcriptome-wide measurement of poly(A) tails and their base composition.
Main Methods:
- Utilized the PacBio HiFi sequencing platform.
- Developed PAIso-seq for poly(A) tail analysis.
- Enabled analysis from nanogram levels of total RNA.
Main Results:
- Accurate measurement of poly(A) tail length and base composition.
- Obtained full-length cDNA sequences alongside poly(A) tail information.
- Enabled comprehensive transcriptome analysis including poly(A) tails.
Conclusions:
- PAIso-seq is a robust method for analyzing poly(A) tails.
- This technique enhances the understanding of mRNA regulation and function.
- PAIso-seq provides valuable insights into transcriptome complexity.
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