Bessière Chloé, Meggetto Fabienne, Gaspin Christine, Fuchs Steffen
Centre de Recherches en Cancérologie de Toulouse (CRCT), UMR-1037 INSERM, UMR-5071 CNRS, Université Toulouse 3, Toulouse, France.
Laboratoire d'Excellence Toulouse Cancer-TOUCAN, Toulouse, France.
Methods Mol Biol. 2025;2880:49-68. doi: 10.1007/978-1-0716-4276-4_3.
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.
环状RNA是一类新兴的非编码RNA，在基因表达调控中发挥着重要作用。虽然存在不同的检测方法，但要揭示其完整序列信息仍较为困难，而这对于理解其功能至关重要。在此，我们描述了一种用于富集环状RNA的方案，这些环状RNA将通过牛津纳米孔长读长测序平台进行长读长测序。这使得能够对环状RNA进行全长测序，并有可能检测到最终的可变剪接事件。我们还通过两种不同工具为生物信息学分析提供了指导，这将检测到高可信度的环状RNA。该方案适用于广泛的疾病背景，将有助于增加对这类RNA的了解。
