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Full-length transcriptome reconstruction reveals a large diversity of RNA and protein isoforms in rat hippocampus.


ABSTRACT: Gene annotation is a critical resource in genomics research. Many computational approaches have been developed to assemble transcriptomes based on high-throughput short-read sequencing, however, only with limited accuracy. Here, we combine next-generation and third-generation sequencing to reconstruct a full-length transcriptome in the rat hippocampus, which is further validated using independent 5´ and 3´-end profiling approaches. In total, we detect 28,268 full-length transcripts (FLTs), covering 6,380 RefSeq genes and 849 unannotated loci. Based on these FLTs, we discover co-occurring alternative RNA processing events. Integrating with polysome profiling and ribosome footprinting data, we predict isoform-specific translational status and reconstruct an open reading frame (ORF)-eome. Notably, a high proportion of the predicted ORFs are validated by mass spectrometry-based proteomics. Moreover, we identify isoforms with subcellular localization pattern in neurons. Collectively, our data advance our knowledge of RNA and protein isoform diversity in the rat brain and provide a rich resource for functional studies.

SUBMITTER: Wang X 

PROVIDER: S-EPMC6825209 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature

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Full-length transcriptome reconstruction reveals a large diversity of RNA and protein isoforms in rat hippocampus.

Wang Xi X   You Xintian X   Langer Julian D JD   Hou Jingyi J   Rupprecht Fiona F   Vlatkovic Irena I   Quedenau Claudia C   Tushev Georgi G   Epstein Irina I   Schaefke Bernhard B   Sun Wei W   Fang Liang L   Li Guipeng G   Hu Yuhui Y   Schuman Erin M EM   Chen Wei W  

Nature communications 20191101 1


Gene annotation is a critical resource in genomics research. Many computational approaches have been developed to assemble transcriptomes based on high-throughput short-read sequencing, however, only with limited accuracy. Here, we combine next-generation and third-generation sequencing to reconstruct a full-length transcriptome in the rat hippocampus, which is further validated using independent 5´ and 3´-end profiling approaches. In total, we detect 28,268 full-length transcripts (FLTs), cover  ...[more]

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