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Transcript assembly improves expression quantification of transposable elements in single-cell RNA-seq data

机译:转录组件改善单细胞RNA-SEQ数据中转换元素的表达量化

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摘要

Transposable elements (TEs) are an integral part of the host transcriptome. TE-containing noncoding RNAs (ncRNAs) show considerable tissue specificity and play important roles during development, including stem cell maintenance and cell differentiation. Recent advances in single-cell RNA-seq (scRNA-seq) revolutionized cell type–specific gene expression analysis. However, effective scRNA-seq quantification tools tailored for TEs are lacking, limiting our ability to dissect TE expression dynamics at single-cell resolution. To address this issue, we established a TE expression quantification pipeline that is compatible with scRNA-seq data generated across multiple technology platforms. We constructed TE-containing ncRNA references using bulk RNA-seq data and showed that quantifying TE expression at the transcript level effectively reduces noise. As proof of principle, we applied this strategy to mouse embryonic stem cells and successfully captured the expression profile of endogenous retroviruses in single cells. We further expanded our analysis to scRNA-seq data from early stages of mouse embryogenesis. Our results illustrated the dynamic TE expression at preimplantation stages and revealed 146 TE-containing ncRNA transcripts with substantial tissue specificity during gastrulation and early organogenesis.
机译:可转换元素(TES)是宿主转录组的组成部分。含TE的非致rNA(NCRNA)显示出相当大的组织特异性并在发育过程中发挥重要作用,包括干细胞维持和细胞分化。单细胞RNA-SEQ(SCRNA-SEQ)彻底的细胞类型特异性基因表达分析的最新进展。然而,缺乏针对TES定制的有效的SCRNA-SEQ定量工具,限制了我们在单细胞分辨率下解析TE表达动态的能力。为了解决这个问题,我们建立了一个TE表达量化管道,它与多个技术平台产生的SCRNA-SEQ数据兼容。我们使用批量RNA-SEQ数据构建含TE的NCRNA引用,并显示在转录物水平上定量TE表达有效地降低了噪声。作为原理的证据,我们将该策略应用于小鼠胚胎干细胞,并成功地捕获了单细胞中内源性逆转录病毒的表达谱。我们进一步扩展了我们对小鼠胚胎发生的早期阶段的ScrNA-SEQ数据的分析。我们的结果说明了预催化阶段的动态TE表达,并揭示了含有大量组织特异性的含有146个TE的NCRNA转录物,并且在脱气期间和早期的器官发生。

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