Full-length transcriptome reconstruction reveals large diversity of RNA isoforms and open reading frames in rat hippocampus

We developed a hybrid-sequencing workflow, combining next-generation and third-generation sequencing, to reconstruct full-length transcriptomes. Integrating with polysome profiling and ribosome footprinting data, we predicted isoform–specific translational status and reconstructed ORFeome. Moreover, we identified isoforms with specific subcellular localization pattern in neurons.本研究开发了一套结合二代测序（next-generation sequencing）与三代测序（third-generation sequencing）的混合测序流程，用于构建全长转录组。结合多聚核糖体谱分析（polysome profiling）与核糖体足迹谱数据（ribosome footprinting），本研究预测了转录本异构体（isoform）特异性的翻译状态，并构建了开放阅读框组（ORFeome）。此外，本研究还在神经元中鉴定出具有特定亚细胞定位模式的转录本异构体。