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Discovery and characterization of new transcripts from RNA-seq data in mouse CD4 + T cells

机译:从小鼠CD4 + T细胞中RNA-seq数据中发现和表征新的转录本

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Despite the routine application of RNA-seq technology to profile cellular transcriptomes and report novel splice variants, the identification and validation of new transcripts remain underexplored. We prepared two RNA-seq libraries from resting and T cell receptor-stimulated mouse CD4 + T cells. Transcripts unknown to Ensembl represent as much as 5% of the assembled transcripts and are robustly expressed but do not show the same degree of evolutionary conservation or exon distribution of known transcripts, or of novel splice isoforms. Here we present a straightforward and generally applicable computational/experimental workflow that we apply to characterise and experimentally validate 23 mouse transcripts from the RNA-seq libraries that were uncharacterised by Ensembl. Of these, 7 are not supported by any transcript database and therefore are likely to encode new messages. Furthermore, we also report the fast up-regulation of important regulatory molecules only 4h post-stimulation of the T cell receptor, which calls for a more detailed investigation into early CD4 + T cell activation mechanisms.
机译:尽管RNA-seq技术的常规应用是分析细胞转录组并报告新的剪接变体,但对新转录本的鉴定和验证仍未得到充分研究。我们从静止和T细胞受体刺激的小鼠CD4 + T细胞制备了两个RNA-seq库。 Ensembl未知的转录本最多可代表组装的转录本的5%,并且表达稳健,但与已知的转录本或新型剪接同工型没有相同程度的进化保守性或外显子分布。在这里,我们介绍了一种简单易用且普遍适用的计算/实验工作流程,我们将其用于表征和实验验证来自Ensembl所不具备的RNA-seq库的23个小鼠转录本。其中的7个不受任何笔录数据库支持,因此很可能会编码新消息。此外,我们还报告了仅在刺激T细胞受体后4小时内重要调节分子的快速上调,这需要对早期CD4 + T细胞激活机制进行更详细的研究。

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