Cargando…

Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo

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-...

Descripción completa

Detalles Bibliográficos
Autores principales: Hoppe, Caroline, Ashe, Hilary L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7982776/
https://www.ncbi.nlm.nih.gov/pubmed/33778778
http://dx.doi.org/10.1016/j.xpro.2021.100379
_version_ 1783667791175352320
author Hoppe, Caroline
Ashe, Hilary L.
author_facet Hoppe, Caroline
Ashe, Hilary L.
author_sort Hoppe, Caroline
collection PubMed
description 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).
format Online
Article
Text
id pubmed-7982776
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-79827762021-03-25 Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo Hoppe, Caroline Ashe, Hilary L. STAR Protoc Protocol 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). Elsevier 2021-03-18 /pmc/articles/PMC7982776/ /pubmed/33778778 http://dx.doi.org/10.1016/j.xpro.2021.100379 Text en © 2021 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Hoppe, Caroline
Ashe, Hilary L.
Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo
title Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo
title_full Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo
title_fullStr Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo
title_full_unstemmed Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo
title_short Live imaging and quantitation of nascent transcription using the MS2/MCP system in the Drosophila embryo
title_sort live imaging and quantitation of nascent transcription using the ms2/mcp system in the drosophila embryo
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7982776/
https://www.ncbi.nlm.nih.gov/pubmed/33778778
http://dx.doi.org/10.1016/j.xpro.2021.100379
work_keys_str_mv AT hoppecaroline liveimagingandquantitationofnascenttranscriptionusingthems2mcpsysteminthedrosophilaembryo
AT ashehilaryl liveimagingandquantitationofnascenttranscriptionusingthems2mcpsysteminthedrosophilaembryo