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Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo

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 Drosophila embryos. These protocols use the SunTag system, in which the protein of interest is visual...

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Autores principales: Vinter, Daisy J., 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/PMC8450298/
https://www.ncbi.nlm.nih.gov/pubmed/34585149
http://dx.doi.org/10.1016/j.xpro.2021.100812
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author Vinter, Daisy J.
Hoppe, Caroline
Ashe, Hilary L.
author_facet Vinter, Daisy J.
Hoppe, Caroline
Ashe, Hilary L.
author_sort Vinter, Daisy J.
collection PubMed
description 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 Drosophila 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).
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spelling pubmed-84502982021-09-27 Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo Vinter, Daisy J. Hoppe, Caroline Ashe, Hilary L. STAR Protoc Protocol 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 Drosophila 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). Elsevier 2021-09-13 /pmc/articles/PMC8450298/ /pubmed/34585149 http://dx.doi.org/10.1016/j.xpro.2021.100812 Text en © 2021 The Authors https://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
Vinter, Daisy J.
Hoppe, Caroline
Ashe, Hilary L.
Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo
title Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo
title_full Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo
title_fullStr Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo
title_full_unstemmed Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo
title_short Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo
title_sort live and fixed imaging of translation sites at single mrna resolution in the drosophila embryo
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8450298/
https://www.ncbi.nlm.nih.gov/pubmed/34585149
http://dx.doi.org/10.1016/j.xpro.2021.100812
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