Custom long non-coding RNA capture enhances detection sensitivity in different human sample types

定制的长链非编码RNA捕获技术可提高不同类型人类样本的检测灵敏度

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作者:Annelien Morlion ,Celine Everaert ,Justine Nuytens ,Eva Hulstaert ,Jo Vandesompele ,Pieter Mestdagh

Abstract

Long non-coding RNAs (lncRNAs) are a heterogeneous group of transcripts that lack protein coding potential and display regulatory functions in various cellular processes. As a result of their cell- and cancer-specific expression patterns, lncRNAs have emerged as potential diagnostic and therapeutic targets. The accurate characterization of lncRNAs in bulk transcriptome data remains challenging due to their low abundance compared to protein coding genes. To tackle this issue, we describe a unique short-read custom lncRNA capture sequencing approach that relies on a comprehensive set of 565,878 capture probes for 49,372 human lncRNA genes. This custom lncRNA capture approach was evaluated on various sample types ranging from artificial high-quality RNA mixtures to more challenging formalin-fixed paraffin-embedded tissue and biofluid material. The custom enrichment approach allows the detection of a more diverse repertoire of lncRNAs, with better reproducibility and higher coverage compared to classic total RNA-sequencing. Keywords: FFPE; RNA abundance; RNA expression; RNA sequencing; biofluid; lncRNA; lncRNome; probes.

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