Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO, USA.
Methods Mol Biol. 2025;2875:99-110. doi: 10.1007/978-1-0716-4248-1_9.
Recent advances in fluorescence microscopy have now made it possible to measure the translation dynamics of individual RNA in living cells and in multiple colors. Here we describe a protocol that exploits these recent advances to simultaneously image the translation of two open reading frames encoded on a single reporter RNA yet frameshifted with respect to each other. This enables precise measurements of frameshifting dynamics and efficiency from specific frameshift stimulatory sequences, all with single-RNA precision.
近年来荧光显微镜技术的进步使得在活细胞中以多种颜色测量单个 RNA 的翻译动力学成为可能。在这里，我们描述了一种利用这些最新进展的方案，该方案能够同时对单个报告 RNA 上编码的两个开放阅读框进行成像，但相对于彼此发生移码。这使得能够从特定的移码刺激序列中精确测量移码动力学和效率，所有这些都具有单个 RNA 的精度。
