Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
Science. 2023 Jun 30;380(6652):eadd3067. doi: 10.1126/science.add3067.
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.
信使 RNA（mRNA）翻译的精确调控是细胞生理转录后基因调控的关键步骤。然而，系统地在转录组水平上以空间和单细胞分辨率研究 mRNA 翻译仍然是一个挑战。在这里，我们报告了核糖体结合的 mRNA 作图（RIBOmap）的开发，这是一种高度多重化的三维原位分析方法，用于检测细胞翻译组。对 HeLa 细胞中的 981 个基因进行 RIBOmap 分析，揭示了细胞周期依赖性的翻译控制和功能基因模块的共翻译。我们对小鼠脑组织中的 5413 个基因进行了作图，为 119173 个细胞生成了空间分辨的单细胞翻译图谱，并揭示了细胞类型特异性和脑区特异性的翻译调控，包括少突胶质细胞成熟过程中的翻译重构。我们的方法在完整脑组织网络中的神经元和神经胶质细胞中检测到广泛的局域翻译模式。
