Single cell quantification of translation control in early mouse development

脊椎动物胚胎发育的最初阶段是在没有转录的情况下进行的。 因此，植入前胚胎的蛋白质丰度受转录后调控。 基因表达程序。特别是，新的蛋白质合成是高产所必需的。 胚胎发生作为翻译的抑制阻止了合子基因组的激活并导致 发展受阻。尽管有这一关键功能，但单个哺乳动物的翻译动力学 由于技术限制，胚胎一直没有被开发出来。要解决这一关键差距并 技术需求，我们建议全面和定量地确定 小鼠单核苷酸和单胚胎发育早期的翻译调控 决议。拟议的研究将应用我的实验室开发的一项变革性技术 能够测量单个植入前胚胎的翻译。在这项研究中，我们将 确定mRNAs的等位基因特异性核糖体结合并表征潜在的 机械装置。这项拟议的研究将具有重要意义，因为它将是第一个全转录范围的研究 单胚胎着床前哺乳动物发育中的翻译控制特征 并将揭示早期胚胎如何执行特定的基因表达程序来塑造 它的蛋白质组。

The initial stages of vertebrate embryo development proceed in the absence of transcription. Consequently, protein abundance of preimplantation embryos is regulated by post-transcriptional gene expression programs. In particular, new protein synthesis is required for productive embryogenesis as inhibition of translation prevents activation of the zygotic genome and leads to arrested development. Despite this critical function, translational dynamics in single mammalian embryos have remained unexplored due to technological limitations. To address this critical gap and technological need, we propose to comprehensively and quantitatively determine the dynamics of translational control during early mouse development at single nucleotide and single embryo resolution. The proposed research will apply a transformative technology that my lab developed enabling the measurement of translation in single preimplantation embryos. In this study, we will determine allele-specific ribosome engagement of mRNAs and characterize the underlying mechanisms. The proposed study will be significant as it will be the first transcriptome-wide characterization of translational control in preimplantation mammalian development at single embryo resolution and will reveal how the early embryo executes specific gene expression programs to shape its proteome.