以单核糖核酸精度对活细胞中核糖体框架转换动态进行成像和定量。

Q4 Biochemistry, Genetics and Molecular Biology Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4248-1_9

Kenneth R Lyon, Tatsuya Morisaki, Timothy J Stasevich

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引用次数: 0

摘要

荧光显微技术的最新进展使测量活细胞中单个 RNA 的多色翻译动态成为可能。在这里，我们描述了一种利用这些最新进展同时对编码在单个报告 RNA 上的两个开放阅读框的翻译进行成像的方案，但这两个阅读框是相互移帧的。这样就能通过特定的移帧刺激序列精确测量移帧动态和效率，而且所有测量都能精确到单条 RNA。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Imaging and Quantifying Ribosomal Frameshifting Dynamics with Single-RNA Precision in Live Cells.

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.

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.