Spatially resolved single-cell translatomics at molecular resolution

Science. 2023 Jun 30;380(6652):eadd3067. doi: 10.1126/science.add3067. Epub 2023 Jun 30.

Abstract

The precise control of messenger RNA (mRNA) translation is a crucial step in posttranscriptional gene regulation of cellular physiology. However, it remains a challenge to systematically study mRNA translation at the transcriptomic scale with spatial and single-cell resolution. Here, we report the development of ribosome-bound mRNA mapping (RIBOmap), a highly multiplexed three-dimensional in situ profiling method to detect cellular translatome. RIBOmap profiling of 981 genes in HeLa cells revealed cell cycle-dependent translational control and colocalized translation of functional gene modules. We mapped 5413 genes in mouse brain tissues, yielding spatially resolved single-cell translatomic profiles for 119,173 cells and revealing cell type-specific and brain region-specific translational regulation, including translation remodeling during oligodendrocyte maturation. Our method detected widespread patterns of localized translation in neuronal and glial cells in intact brain tissue networks.

MeSH terms

  • Animals
  • Brain* / metabolism
  • Chromosome Mapping* / methods
  • Gene Expression Regulation
  • HeLa Cells
  • Humans
  • Mice
  • Neuroglia* / metabolism
  • Neurons* / metabolism
  • Protein Biosynthesis*
  • RNA, Messenger* / genetics
  • RNA, Messenger* / metabolism
  • Single-Cell Gene Expression Analysis* / methods

Substances

  • RNA, Messenger