Single-cell mRNA isoform diversity in the mouse brain.
Single-cell mRNA isoform diversity in the mouse brain.
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DOI：
10.1186/s12864-017-3528-6
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发表时间：
2017-02-03
期刊：
影响因子：
4.4
通讯作者：
Linnarsson S
中科院分区：
文献类型：
--
作者：
Karlsson K;Linnarsson S
替代mRNA同种型的使用是哺乳动物细胞中蛋白质多样性的重要来源。这种现象已经在大量组织中得到了广泛的研究，然而，目前还不清楚这种多样性如何反映在单细胞中。在这里，我们使用长读段测序技术结合独特的分子标识符（UMI），以揭示模式的替代全长亚型表达的单细胞从小鼠大脑。我们发现了令人惊讶的数量的异构体多样性，即使在应用保守的定义是什么构成了异构体。基因往往有一个或几个亚型高度表达和大量的亚型表达水平较低。然而，对于许多基因来说，几乎每个测序的mRNA分子都是独特的，许多事件影响了编码区，这表明单细胞中以前未知的蛋白质多样性。编码区的外显子连接比非编码区的外显子连接更不容易发生剪接错误，表明剪接供体和受体效率上的纯化选择。我们的研究结果表明，mRNA异构体的多样性是一个重要来源的生物变异性也在单细胞。本文的在线版本（doi：10.1186/s12864-017-3528-6）包含补充材料，可供授权用户使用。
Alternative mRNA isoform usage is an important source of protein diversity in mammalian cells. This phenomenon has been extensively studied in bulk tissues, however, it remains unclear how this diversity is reflected in single cells. Here we use long-read sequencing technology combined with unique molecular identifiers (UMIs) to reveal patterns of alternative full-length isoform expression in single cells from the mouse brain. We found a surprising amount of isoform diversity, even after applying a conservative definition of what constitutes an isoform. Genes tend to have one or a few isoforms highly expressed and a larger number of isoforms expressed at a low level. However, for many genes, nearly every sequenced mRNA molecule was unique, and many events affected coding regions suggesting previously unknown protein diversity in single cells. Exon junctions in coding regions were less prone to splicing errors than those in non-coding regions, indicating purifying selection on splice donor and acceptor efficiency. Our findings indicate that mRNA isoform diversity is an important source of biological variability also in single cells. The online version of this article (doi:10.1186/s12864-017-3528-6) contains supplementary material, which is available to authorized users.