Faculty of Biology, Medicine and Health, University of Manchester, Manchester M13 9PT, UK.
STAR Protoc. 2021 Sep 13;2(3):100812. doi: 10.1016/j.xpro.2021.100812. eCollection 2021 Sep 17.
Significant regulation of gene expression is mediated at the translation level. Here, we describe protocols for imaging and analysis of translation at single mRNA resolution in both fixed and living embryos. These protocols use the SunTag system, in which the protein of interest is visualized by inserting a peptide array that is recognized by a single chain antibody. Simultaneous detection of individual mRNAs using the MS2/MCP system or by smFISH allows translation sites to be identified and quantified. For complete information on the generation and use of this protocol, please refer to Vinter et al. (2021).
基因表达的重要调控是在翻译水平上进行的。在这里，我们描述了在固定和活体胚胎中以单个 mRNA 分辨率进行翻译的成像和分析的方案。这些方案使用 SunTag 系统，其中通过插入被单个链抗体识别的肽阵列来可视化感兴趣的蛋白质。使用 MS2/MCP 系统或 smFISH 同时检测单个 mRNA，可识别和定量翻译位点。有关此方案的生成和使用的完整信息，请参阅 Vinter 等人。（2021）。
