美国德克萨斯大学Can Cenik研究组实现小鼠早期发育过程中核糖体结合的单细胞定量分析。2023年6月21日，《自然》杂志在线发表了这项成果。

研究人员表示，翻译调节对哺乳动物早期胚胎发育至关重要。然而，以前的研究仅限于批量测量，排除了对翻译调节的精确测定，包括等位基因特异性分析。

为了应对这一挑战，研究人员开发了一种新的微流控等离子泳（ITP）方法，命名为RIBOsome profiling via ITP（Ribo-ITP），并对小鼠早期发育过程中单个卵母细胞和胚胎中的翻译进行了描述。研究人员发现不同的翻译效率是调节参与中心体组织和RNA的N6-甲基腺苷修饰基因的一个关键机制。这个高覆盖率测量使得研究人员首次分析了早期发育中的等位基因特异性核糖体参与。这导致了阶段性地发现了子代RNA与核糖体的不同接触，以及表现出等位基因偏向表达转录本的翻译效率降低。

通过整合这些测量和蛋白质组学数据，研究人员发现生发泡阶段的卵母细胞中的核糖体占有率是决定子宫内蛋白质丰度的主要因素。Ribo-ITP方法将通过从包括单细胞在内的超低输入样品中提供高覆盖率和高分辨率的核糖体占有率测量，从而实现众多的应用。

附：英文原文

Title: Single-cell quantification of ribosome occupancy in early mouse development

Author: Ozadam, Hakan, Tonn, Tori, Han, Crystal M., Segura, Alia, Hoskins, Ian, Rao, Shilpa, Ghatpande, Vighnesh, Tran, Duc, Catoe, David, Salit, Marc, Cenik, Can

Issue&Volume: 2023-06-21

Abstract: Translation regulation is critical for early mammalian embryonic development1. However, previous studies had been restricted to bulk measurements2, precluding precise determination of translation regulation including allele-specific analyses. Here, to address this challenge, we developed a novel microfluidic isotachophoresis (ITP) approach, named RIBOsome profiling via ITP (Ribo-ITP), and characterized translation in single oocytes and embryos during early mouse development. We identified differential translation efficiency as a key mechanism regulating genes involved in centrosome organization and N6-methyladenosine modification of RNAs. Our high-coverage measurements enabled, to our knowledge, the first analysis of allele-specific ribosome engagement in early development. These led to the discovery of stage-specific differential engagement of zygotic RNAs with ribosomes and reduced translation efficiency of transcripts exhibiting allele-biased expression. By integrating our measurements with proteomics data, we discovered that ribosome occupancy in germinal vesicle-stage oocytes is the predominant determinant of protein abundance in the zygote. The Ribo-ITP approach will enable numerous applications by providing high-coverage and high-resolution ribosome occupancy measurements from ultra-low input samples including single cells.

DOI: 10.1038/s41586-023-06228-9

Source: https://www.nature.com/articles/s41586-023-06228-9