Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
Methods Mol Biol. 2021;2252:189-200. doi: 10.1007/978-1-0716-1150-0_8.
Modern methods of genome editing enable the rapid generation of mouse models to study the regulation of protein synthesis. At the same time, few options are available to study translation in rodents as the animal's complexity severely limits the repertoire of experimental tools. Here we describe a method to monitor translation in mice and other small animals. The technique is based on a ribosome profiling and specifically tailored toward measuring translation elongation. However, it can be easily applied for short upstream reading frames discovery. The advantage of this method is the ability to study translation in fully developed animals without extracting and subculturing cells, therefore, maintaining unperturbed physiological conditions.
现代的基因组编辑方法使快速生成研究蛋白质合成调控的小鼠模型成为可能。与此同时,由于动物的复杂性严重限制了实验工具的种类,研究啮齿动物中的翻译可用的方法很少。在这里,我们描述了一种在小鼠和其他小型动物中监测翻译的方法。该技术基于核糖体图谱分析,并专门针对测量翻译延伸进行了优化。然而,它也可以很容易地应用于短的上游阅读框的发现。这种方法的优点是能够在完全发育的动物中研究翻译,而无需提取和传代细胞,因此可以保持未受干扰的生理条件。