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Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes

Rahman, Raza Ur
Ahmad, Iftikhar
Li, Zixiu
Sparks, Robert P
Ben Saad, Amel
Mullen, Alan C
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Abstract

Single cell RNA sequencing (scRNA-seq) has revolutionized the study of gene expression in individual cell types, but scRNA-seq studies have focused primarily on expression of protein-coding genes. Long noncoding RNAs (lncRNAs) are more diverse than protein-coding genes, yet remain underexplored in part because they are underrepresented in reference annotations applied to scRNA-seq. Merging annotations containing protein-coding and lncRNA genes is not sufficient, because the addition of lncRNA genes that overlap in sense and antisense with protein-coding genes will affect how reads are counted for both protein-coding and lncRNA genes. Here, we introduce Singletrome, a Singularity image that integrates protein-coding and lncRNA gene transfer format (GTF) annotations to generate enhanced annotations that take into account the sense and antisense overlap of annotated genes, maps scRNA-seq data, and produces files for downstream analysis and visualization. With Singletrome, we detected thousands of lncRNAs not included in GENCODE, clustered cell types based solely on lncRNA expression, and demonstrated that machine learning can predict cell type and disease through lncRNAs alone. This comprehensive annotation will allow mapping of lncRNA expression across cell types of the human body, facilitating the development of an atlas of human lncRNAs in health and disease with the ability to integrate new lncRNA annotations as they become available.

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Rahman RU, Ahmad I, Li Z, Sparks RP, Ben Saad A, Mullen AC. Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes. Sci Rep. 2025 Aug 12;15(1):29542. doi: 10.1038/s41598-025-13528-9. PMID: 40796606; PMCID: PMC12344142.

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10.1038/s41598-025-13528-9
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40796606
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Open Access: This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit https://creativecommons.org/licenses/by-nc-nd/4.0/. © The Author(s) 2025