ArticleRNA biology2021
5´XP sRNA-seq: efficient identification of transcripts with and without 5´ phosphorylation reveals evolutionary conserved small RNA.
Article in RNA biology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- A method of comprehensive sequencing analysis of the small RNA fragmentome (RiboMarker).RNA (New York, N.Y.) · 2026Article
- Optimized identification and characterization of small RNAs with PANDORA-seq.Nature protocols · 2025Review
- Small RNA modifications: regulatory molecules and potential applications.Journal of hematology & oncology · 2023Review
- Seqpac: a framework for sRNA-seq analysis in R using sequence-based counts.Bioinformatics (Oxford, England) · 2023Article
- Exploring the expanding universe of small RNAs.Nature cell biology · 2022Review
- Cell type-specific analysis by single-cell profiling identifies a stable mammalian tRNA-mRNA interface and increased translation efficiency in neurons.Genome research · 2022Article
- Deciphering the RNA universe in sperm in its role as a vertical information carrier.Environmental epigenetics · 2022Review
- Origins and evolving functionalities of tRNA-derived small RNAs.Trends in biochemical sciences · 2021Review
- Small RNA-Sequencing: Approaches and Considerations for miRNA Analysis.Diagnostics (Basel, Switzerland) · 2021Review
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Small RNA (sRNA) sequencing has been critical for our understanding of many cellular processes, including gene regulation. Nonetheless, the varying biochemical properties of sRNA, such as 5´ nucleotide modifications, make many sRNA subspecies incompatible with common protocols for sRNA sequencing. Here we describe 5XP-seq that outlines a novel strategy that captures a more complete picture of sRNA. By tagging 5´P sRNA during library preparation, 5XP-seq combines an open approach that includes all types of 5'-terminal modifications (5´X), with a selective approach for 5-phosphorylated sRNA (5´P). We show that 5XP-seq not only enriches phosphorylated miRNA and piRNA but successfully discriminates these sRNA from all other sRNA species. We further demonstrate the importance of this strategy by successful inter-species validation of sRNAs that would have otherwise failed, including human to insect translation of several tRNA (tRFs) and rRNA (rRFs) fragments. By combining 5´ insensitive library strategies with 5´ sensitive tagging, we have successfully tackled an intrinsic bias in modern sRNA sequencing that will help us reveal the true complexity and the evolutionary significance of the sRNA world.
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Registered trials
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