Publication Date
6-29-2023
Document Type
Article
Publication Title
Nature
Volume
618
Issue
7967
DOI
10.1038/s41586-023-06228-9
First Page
1057
Last Page
1064
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 N 6-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.
Funding Number
CA204522
Funding Sponsor
National Institutes of Health
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
Department
Mechanical Engineering
Recommended Citation
Hakan Ozadam, Tori Tonn, Crystal M. Han, Alia Segura, Ian Hoskins, Shilpa Rao, Vighnesh Ghatpande, Duc Tran, David Catoe, Marc Salit, and Can Cenik. "Single-cell quantification of ribosome occupancy in early mouse development" Nature (2023): 1057-1064. https://doi.org/10.1038/s41586-023-06228-9
