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Circular RNAs are an emerging class of noncoding RNAs playing important functions in gene expression regulation. While different detection methods exist, the information of their full sequence remains more difficult to reveal, but is crucial to understand their function. Here, we describe a protocol to enrich for circRNAs that are subjected to long-read sequencing with the Oxford Nanopore long-read sequencing platform. This allows to sequence circRNAs in full length and gives the possibility to detect eventual alternative splicing events. We further provide guidance for the bioinformatics analysis by two different tools, which will detect circRNAs of high confidence. This protocol is applicable to a wide range of disease contexts and will help to increase knowledge about this RNA class.

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Identification of Circular RNA Variants by Oxford Nanopore Long-Read Sequencing

  • 实验方案

  • 发表日期：

    2025年

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  • Chloé Bessière,
  • Fabienne Meggetto,
  • Christine Gaspin,
  • Steffen Fuchs

摘要

Circular RNAs are an emerging class of noncoding RNAs playing important functions in gene expression regulation. While different detection methods exist, the information of their full sequence remains more difficult to reveal, but is crucial to understand their function. Here, we describe a protocol to enrich for circRNAs that are subjected to long-read sequencing with the Oxford Nanopore long-read sequencing platform. This allows to sequence circRNAs in full length and gives the possibility to detect eventual alternative splicing events. We further provide guidance for the bioinformatics analysis by two different tools, which will detect circRNAs of high confidence. This protocol is applicable to a wide range of disease contexts and will help to increase knowledge about this RNA class.