Recursive splicing and mRNA polyadenylation regulatory circuits govern homeostasis and cell cycle potency of pluripotent cells.

递归剪接和 mRNA 多腺苷酸化调节电路控制多能细胞的稳态和细胞周期效力。

• 批准号: 313408820
• 负责人: Professor Dr. Argyris Papantonis
• 金额: --
• 依托单位: Institut für Pathologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/313408820?language=en
• 关键词: Recursive; splicing; mRNA; polyadenylation; regulatory

Mammalian transcriptomes are extremely complex, with large amounts of RNA copied from non-coding regions, like introns that occupy most of the length of protein-coding genes and are now understood to contribute to gene expression regulation. Our lab was of the first to uncover the existence of recursive splicing in human cells, an elegant mechanism for the stepwise processing of long introns. However, despite significant recent advances, including our work in the first funding round of this Priority Program, many aspects of recursive splicing remain unknown, and are the focal point of this proposal. We need to decipher how specific RS sites are actually selected in the production of mature mRNAs, and we need to dissect the contribution of recursive splicing in cell homeostasis and disease. For this proposal, we present preliminary data, generated within the SPP1935, uncovering a link between recursive splicing sites and disease manifestation across human cell types, as well as results highlighting recursive splicing regulatory circuits in human pluripotent stem cells that are governed by RNPs involving mRNAs and variant U1 transcripts. Thus, we now aim at dissecting the mRNP code behind these circuits by combining genomics, biochemical, and CRISPR/Cas9 genome editing approaches with in vitro differentiation of human pluripotent cells intro different lineages. We see this as a prerequisite for understanding the impact of recursive splicing in cell homeostasis and development, as well as in the revaluation of the functional partition of nuclear space as regards the transcription cycle of human genes. We are convinced that this novel layer of gene expression regulation presents a entryway into deciphering the complexity of mammalian mRNA processing and has implications for both development and disease.

哺乳动物转录组非常复杂，有大量的RNA从非编码区复制，如内含子，占据了蛋白质编码基因的大部分长度，现在被认为有助于基因表达调控。我们的实验室是第一个发现人类细胞中存在递归剪接的实验室，这是一种逐步处理长内含子的优雅机制。然而，尽管最近取得了重大进展，包括我们在该优先计划的第一轮融资中的工作，但递归拼接的许多方面仍然未知，并且是该提案的焦点。我们需要破译在成熟mRNA的产生过程中，特定的RS位点是如何被选择的，我们需要剖析递归剪接在细胞稳态和疾病中的作用。对于这一建议，我们提出了初步的数据，在SPP 1935中产生，揭示了递归剪接位点和疾病表现之间的联系，在人类细胞类型，以及结果突出递归剪接调节回路在人类多能干细胞，由RNP涉及mRNA和变异U1转录。因此，我们现在的目标是通过将基因组学、生物化学和CRISPR/Cas9基因组编辑方法与引入不同谱系的人多能细胞的体外分化相结合，来剖析这些回路背后的mRNP代码。我们认为这是理解递归剪接在细胞稳态和发育中的影响，以及重新评估人类基因转录周期中核空间功能分区的先决条件。我们相信，这一新的基因表达调控层为破译哺乳动物mRNA加工的复杂性提供了一个入口，并对发育和疾病都有影响。