Unveiling Tissue‐Specific RNA Landscapes in Mouse Organs During Fasting and Feeding Using Nanopore Direct RNA Sequencing

Abstract Understanding tissue‐specific RNA landscapes is essential for uncovering the functional mechanisms of key organs in mammals. However, current knowledge remains limited, as short‐read RNA sequencing—the predominant method for assessing gene expression—depends on incomplete gene annotations a...

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Main Authors: Chengfei Jiang, Ping Li, Haiming Cao
Format: Article
Language:English
Published: Wiley 2025-02-01
Series:Advanced Science
Subjects:
Online Access:https://doi.org/10.1002/advs.202408054
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author Chengfei Jiang
Ping Li
Haiming Cao
author_facet Chengfei Jiang
Ping Li
Haiming Cao
author_sort Chengfei Jiang
collection DOAJ
description Abstract Understanding tissue‐specific RNA landscapes is essential for uncovering the functional mechanisms of key organs in mammals. However, current knowledge remains limited, as short‐read RNA sequencing—the predominant method for assessing gene expression—depends on incomplete gene annotations and struggles to resolve the diverse transcripts produced by genes. To address these limitations, an integrative approach combining nanopore direct RNA sequencing (DRS), ATAC‐Seq, and short‐read RNA‐seq is used. This method enabled the analysis of RNA landscapes across major mouse organs under fasting and fed conditions, representing two extremes of the caloric cycle. This study uncovered tens of thousands of novel transcripts and identified hundreds of genes with tissue‐specific expression, revealing additional layers of regulated pathways within each organ that conventional short‐read RNA‐seq cannot resolve. By profiling transcript expression across multiple organs under identical conditions, it is conducted comparative analyses exposing significant differences in transcript isoforms and regulations. Moreover, nanopore DRS revealed dynamic changes in poly(A) tail length and m6A modifications of transcripts, many regulated in a tissue‐specific manner. These changes likely contribute to functional differentiation and metabolic specialization of various organs. Collectively, this findings reveal previously unrecognized layers of gene regulation, offering new insights into the metabolic basis of organ function.
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spelling doaj-art-fcbd1c56c9e54bab80da75d43c77980d2025-02-04T13:14:55ZengWileyAdvanced Science2198-38442025-02-01125n/an/a10.1002/advs.202408054Unveiling Tissue‐Specific RNA Landscapes in Mouse Organs During Fasting and Feeding Using Nanopore Direct RNA SequencingChengfei Jiang0Ping Li1Haiming Cao2Cardiovascular Branch, National Heart, Lung and Blood Institute National Institutes of Health Bethesda MD 20892 USACardiovascular Branch, National Heart, Lung and Blood Institute National Institutes of Health Bethesda MD 20892 USACardiovascular Branch, National Heart, Lung and Blood Institute National Institutes of Health Bethesda MD 20892 USAAbstract Understanding tissue‐specific RNA landscapes is essential for uncovering the functional mechanisms of key organs in mammals. However, current knowledge remains limited, as short‐read RNA sequencing—the predominant method for assessing gene expression—depends on incomplete gene annotations and struggles to resolve the diverse transcripts produced by genes. To address these limitations, an integrative approach combining nanopore direct RNA sequencing (DRS), ATAC‐Seq, and short‐read RNA‐seq is used. This method enabled the analysis of RNA landscapes across major mouse organs under fasting and fed conditions, representing two extremes of the caloric cycle. This study uncovered tens of thousands of novel transcripts and identified hundreds of genes with tissue‐specific expression, revealing additional layers of regulated pathways within each organ that conventional short‐read RNA‐seq cannot resolve. By profiling transcript expression across multiple organs under identical conditions, it is conducted comparative analyses exposing significant differences in transcript isoforms and regulations. Moreover, nanopore DRS revealed dynamic changes in poly(A) tail length and m6A modifications of transcripts, many regulated in a tissue‐specific manner. These changes likely contribute to functional differentiation and metabolic specialization of various organs. Collectively, this findings reveal previously unrecognized layers of gene regulation, offering new insights into the metabolic basis of organ function.https://doi.org/10.1002/advs.202408054ATAC‐Seqenergy metabolismnanopore direct RNA sequencingpoly(A) tail lengthRNA m6A modification
spellingShingle Chengfei Jiang
Ping Li
Haiming Cao
Unveiling Tissue‐Specific RNA Landscapes in Mouse Organs During Fasting and Feeding Using Nanopore Direct RNA Sequencing
Advanced Science
ATAC‐Seq
energy metabolism
nanopore direct RNA sequencing
poly(A) tail length
RNA m6A modification
title Unveiling Tissue‐Specific RNA Landscapes in Mouse Organs During Fasting and Feeding Using Nanopore Direct RNA Sequencing
title_full Unveiling Tissue‐Specific RNA Landscapes in Mouse Organs During Fasting and Feeding Using Nanopore Direct RNA Sequencing
title_fullStr Unveiling Tissue‐Specific RNA Landscapes in Mouse Organs During Fasting and Feeding Using Nanopore Direct RNA Sequencing
title_full_unstemmed Unveiling Tissue‐Specific RNA Landscapes in Mouse Organs During Fasting and Feeding Using Nanopore Direct RNA Sequencing
title_short Unveiling Tissue‐Specific RNA Landscapes in Mouse Organs During Fasting and Feeding Using Nanopore Direct RNA Sequencing
title_sort unveiling tissue specific rna landscapes in mouse organs during fasting and feeding using nanopore direct rna sequencing
topic ATAC‐Seq
energy metabolism
nanopore direct RNA sequencing
poly(A) tail length
RNA m6A modification
url https://doi.org/10.1002/advs.202408054
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AT pingli unveilingtissuespecificrnalandscapesinmouseorgansduringfastingandfeedingusingnanoporedirectrnasequencing
AT haimingcao unveilingtissuespecificrnalandscapesinmouseorgansduringfastingandfeedingusingnanoporedirectrnasequencing