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Diversification of stem cell molecular repertoire by alternative splicing

机译：通过选择性剪接使干细胞分子库多样化

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摘要

Complete information regarding transcriptional and posttranscriptional gene regulation in stem cells is necessary to understand the regulation of self-renewal and differentiation. Alternative splicing is a prevalent mode of posttranscriptional regulation, and occurs in approximately one half of all mammalian genes. The frequency and functional impact of alternative splicing in stem cells are yet to be determined. In this study we combine computational and experimental methods to identify splice variants in embryonic and hematopoietic stem cells on a genome-wide scale. Using EST collections derived from stem cells, we detect alternative splicing in > 1,000 genes. Systematic RT-PCR and sequencing studies show confirmation of computational predictions at a level of 80%. We find that alternative splicing can modify multiple components of signaling pathways important for stem cell function. We also analyze the distribution of splice variants across different classes of genes. We find that tissue-specific genes have a higher tendency to undergo alternative splicing than ubiquitously expressed genes. Furthermore, the patterns of alternative splicing are only weakly conserved between orthologous genes in human and mouse. Our studies reveal extensive modification of the stem cell molecular repertoire by alternative splicing and provide insights into its overall role as a mechanism of generating genomic diversity.
机译：关于干细胞转录和转录后基因调控的完整信息对于理解自我更新和分化的调控是必需的。选择性剪接是转录后调控的一种普遍模式，发生在所有哺乳动物基因的大约一半中。干细胞中选择性剪接的频率和功能影响尚待确定。在这项研究中，我们结合了计算和实验方法，以鉴定全基因组范围内的胚胎和造血干细胞中的剪接变体。使用源自干细胞的EST集合，我们可以检测到> 1,000个基因中的其他剪接。系统的RT-PCR和测序研究表明计算预测的确认率为80％。我们发现替代剪接可以修饰对干细胞功能重要的信号通路的多个组成部分。我们还分析了不同基因类别之间剪接变体的分布。我们发现组织特异性基因比无处不在表达的基因具有更高的发生选择性剪接的趋势。此外，在人和小鼠的直系同源基因之间，选择性剪接的模式仅是弱保守的。我们的研究揭示了通过选择性剪接对干细胞分子库进行的广泛修饰，并深入了解了其作为产生基因组多样性的机制的总体作用。

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