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Updated: Oct 31, 2025
12:01
3' End Sequencing Library Preparation with A-seq2
Published on: October 10, 2017
10.8K
TRENDseq-A highly multiplexed high throughput RNA 3' end sequencing for mapping alternative polyadenylation.
Anton Ogorodnikov1, Sven Danckwardt2
1Posttranscriptional Gene Regulation, University Medical Centre Mainz, Mainz, Germany; Institute for Clinical Chemistry and Laboratory Medicine, University Medical Centre Mainz, Mainz, Germany; Centre for Thrombosis and Hemostasis (CTH), University Medical Centre Mainz, Mainz, Germany.
Methods in Enzymology
|June 29, 2021
Summary
Alternative polyadenylation (APA), a key gene regulator affecting over 70% of genes, generates diverse mRNA isoforms. We present TRENDseq, a sensitive method for transcriptome-wide APA profiling, crucial for understanding disease mechanisms.
Area of Science:
- Molecular Biology
- Genetics
- Transcriptomics
Background:
- Alternative polyadenylation (APA) significantly impacts gene regulation, influencing over 70% of genes.
- APA generates transcript isoforms with distinct 3' ends, expanding transcriptome diversity and affecting protein output, localization, and function.
- Dysregulation of APA is linked to various disorders, highlighting the need for robust methods to study its landscape.
Purpose of the Study:
- To provide a detailed protocol for TRENDseq, a method for transcriptome-wide high-throughput sequencing of polyadenylated RNA 3' ends.
- To enable highly multiplexed and sensitive analysis of APA.
- To facilitate the study of APA in diverse experimental settings, including those with limited sample input.
Main Methods:
- TRENDseq utilizes linear amplification to enhance sensitivity.
- The method allows for highly multiplexed sequencing of RNA 3' ends.
- It requires significantly reduced input material compared to other methods.
Main Results:
- TRENDseq offers improved sensitivity for detecting APA events.
- The protocol enables reproducible and highly multiplexed analysis.
- It is suitable for studying APA in limited biological samples.
Conclusions:
- TRENDseq is a powerful tool for comprehensive APA profiling.
- This method aids in deciphering APA's role in biological processes and disease.
- It supports diagnostic and therapeutic investigations related to APA dysregulation.
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