胚胎中使用 MS2/MCP 系统进行新生转录的实时成像和定量。

Faculty of Biology, Medicine and Health, University of Manchester, Manchester M13 9PT, UK.

STAR Protoc. 2021 Mar 18;2(1):100379. doi: 10.1016/j.xpro.2021.100379. eCollection 2021 Mar 19.

Visualizing transcription live in is providing important new insights into the spatiotemporal regulation of transcription. Here, we describe a protocol to visualize and quantitate transcription from gene loci that are tagged with MS2 stem-loop sequences in the embryo. MS2 stem-loop sequences are recognized by a coat protein fused to a fluorescent protein and visualized with microscopy. We also describe an analysis pipeline to extract and subsequently quantify transcription dynamics. For complete details on the use and execution of this protocol, please refer to Hoppe et al. (2020).

实时可视化转录为转录的时空调控提供了重要的新见解。在这里，我们描述了一种在带有 MS2 茎环序列的胚胎中可视化和定量基因座转录的方案。MS2 茎环序列被融合到荧光蛋白上的外壳蛋白识别，并通过显微镜观察。我们还描述了一种提取和随后量化转录动力学的分析管道。有关该协议的使用和执行的完整详细信息，请参阅 Hoppe 等人。（2020 年）。