Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes

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 underexpl...

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Main Authors: Raza Ur Rahman, Iftikhar Ahmad, Zixiu Li, Robert P Sparks, Amel Ben Saad, Alan C Mullen
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-13528-9
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author Raza Ur Rahman
Iftikhar Ahmad
Zixiu Li
Robert P Sparks
Amel Ben Saad
Alan C Mullen
author_facet Raza Ur Rahman
Iftikhar Ahmad
Zixiu Li
Robert P Sparks
Amel Ben Saad
Alan C Mullen
author_sort Raza Ur Rahman
collection DOAJ
description 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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publishDate 2025-08-01
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spelling doaj-art-fe0b4bfd7e374fdea94232db1a90e50e2025-08-20T03:45:52ZengNature PortfolioScientific Reports2045-23222025-08-0115111610.1038/s41598-025-13528-9Singletrome enhances detection of long noncoding RNAs in single cell transcriptomesRaza Ur Rahman0Iftikhar Ahmad1Zixiu Li2Robert P Sparks3Amel Ben Saad4Alan C Mullen5Division of Gastroenterology, University of Massachusetts Chan Medical SchoolDepartment of Software Engineering, University of Europe for Applied SciencesPopulation and Quantitative Health Sciences, University of Massachusetts Chan Medical SchoolDivision of Gastroenterology, University of Massachusetts Chan Medical SchoolDivision of Gastroenterology, University of Massachusetts Chan Medical SchoolDivision of Gastroenterology, University of Massachusetts Chan Medical SchoolAbstract 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.https://doi.org/10.1038/s41598-025-13528-9
spellingShingle Raza Ur Rahman
Iftikhar Ahmad
Zixiu Li
Robert P Sparks
Amel Ben Saad
Alan C Mullen
Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes
Scientific Reports
title Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes
title_full Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes
title_fullStr Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes
title_full_unstemmed Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes
title_short Singletrome enhances detection of long noncoding RNAs in single cell transcriptomes
title_sort singletrome enhances detection of long noncoding rnas in single cell transcriptomes
url https://doi.org/10.1038/s41598-025-13528-9
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