Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo
Caroline Hoppe, Hilary L. Ashe
Abstract
Visualizing transcription live in Drosophila 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 Drosophila 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).
Topics & Concepts
BiologyTranscription (linguistics)EmbryoTranscription factorComputational biologyGeneCell biologyGeneticsPhilosophyLinguisticsCRISPR and Genetic EngineeringGenomics and Chromatin DynamicsSingle-cell and spatial transcriptomics