Wang Qiuyi, Mao Yuanhui
Bone Marrow Transplantation Center, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
Liangzhu Laboratory, School of Medicine, Zhejiang University, Hangzhou, China.
Adv Biotechnol (Singap). 2023 Dec 1;1(4):6. doi: 10.1007/s44307-023-00006-4.
Ribosome profiling has revolutionized our understanding of gene expression regulation by providing a snapshot of global translation in vivo. This powerful technique enables the investigation of the dynamics of translation initiation, elongation, and termination, and has provided insights into the regulation of protein synthesis under various conditions. Despite its widespread adoption, challenges persist in obtaining high-quality ribosome profiling data. In this review, we discuss the fundamental principles of ribosome profiling and related methodologies, including selective ribosome profiling and translation complex profiling. We also delve into quality control to assess the reliability of ribosome profiling datasets, and the efforts to improve data quality by modifying the standard procedures. Additionally, we highlight recent advancements in ribosome profiling that enable the transition from bulk to low-input and single-cell applications. Single-cell ribosome profiling has emerged as a crucial tool for exploring translation heterogeneity within specific cell populations. However, the challenges of capturing mRNAs efficiently and the sparse nature of footprint reads in single-cell ribosome profiling present ongoing obstacles. The need to refine ribosome profiling techniques remains, especially when used at the single-cell level.
核糖体谱分析通过提供体内全局翻译的快照，彻底改变了我们对基因表达调控的理解。这项强大的技术能够研究翻译起始、延伸和终止的动态过程，并为各种条件下蛋白质合成的调控提供了见解。尽管它已被广泛应用，但在获得高质量核糖体谱分析数据方面仍然存在挑战。在这篇综述中，我们讨论了核糖体谱分析的基本原理和相关方法，包括选择性核糖体谱分析和翻译复合体谱分析。我们还深入探讨了质量控制，以评估核糖体谱分析数据集的可靠性，以及通过修改标准程序来提高数据质量的努力。此外，我们强调了核糖体谱分析的最新进展，这些进展使得从大量样本应用向低输入量和单细胞应用的转变成为可能。单细胞核糖体谱分析已成为探索特定细胞群体内翻译异质性的关键工具。然而，在单细胞核糖体谱分析中有效捕获mRNA的挑战以及足迹读数的稀疏性仍然是持续存在的障碍。改进核糖体谱分析技术的需求仍然存在，特别是在单细胞水平上使用时。
