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

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.