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Current short-read RNA sequencing techniques capture only a limited picture of the human transcriptome, missing novel transcripts and genes that are not in reference annotations. Additionally, conditions that may reveal novel transcription products are often inaccessible using donor human tissues. We describe a protocol that addresses these shortcomings using liver-specific humanized mice and nanopore direct RNA sequencing (DRS). Our protocol captures transcript level changes of human liver transcriptomes under metabolic conditions that cannot be readily studied from human tissues.

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Establishing a De Novo Annotation of Human Liver Transcriptome Based on Long-Read Direct RNA Sequencing Technology and a Liver-Specific Humanized Mouse Model

  • 实验方案

  • 发表日期：

    2025年

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  • Chengfei Jiang,
  • Hang Sun,
  • Haiming Cao

摘要

Current short-read RNA sequencing techniques capture only a limited picture of the human transcriptome, missing novel transcripts and genes that are not in reference annotations. Additionally, conditions that may reveal novel transcription products are often inaccessible using donor human tissues. We describe a protocol that addresses these shortcomings using liver-specific humanized mice and nanopore direct RNA sequencing (DRS). Our protocol captures transcript level changes of human liver transcriptomes under metabolic conditions that cannot be readily studied from human tissues.