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

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

Summary: 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...

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Bibliographic Details
Main Authors: Daisy J. Vinter, Caroline Hoppe, Hilary L. Ashe
Format: Article
Language:English
Published: Elsevier 2021-09-01
Series:STAR Protocols
Subjects:
Cell Biology
Developmental biology
Gene Expression
In Situ Hybridization
Microscopy
Model Organisms
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721005189
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spelling doaj-58675458936343888e7262d75d25ccd82021-09-19T05:01:26ZengElsevierSTAR Protocols2666-16672021-09-0123100812Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryoDaisy J. Vinter0Caroline Hoppe1Hilary L. Ashe2Faculty of Biology, Medicine and Health, University of Manchester, Manchester M13 9PT, UK; Corresponding authorFaculty of Biology, Medicine and Health, University of Manchester, Manchester M13 9PT, UKFaculty of Biology, Medicine and Health, University of Manchester, Manchester M13 9PT, UK; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166721005189Cell BiologyDevelopmental biologyGene ExpressionIn Situ HybridizationMicroscopyModel Organisms
collection DOAJ
language English
format Article
sources DOAJ
author Daisy J. Vinter
Caroline Hoppe
Hilary L. Ashe
spellingShingle Daisy J. Vinter
Caroline Hoppe
Hilary L. Ashe
Live and fixed imaging of translation sites at single mRNA resolution in the Drosophila embryo
STAR Protocols
Cell Biology
Developmental biology
Gene Expression
In Situ Hybridization
Microscopy
Model Organisms
author_facet Daisy J. Vinter
Caroline Hoppe
Hilary L. Ashe
author_sort Daisy J. Vinter
title 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_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_sort live and fixed imaging of translation sites at single mrna resolution in the drosophila embryo
publisher Elsevier
series STAR Protocols
issn 2666-1667
publishDate 2021-09-01
description Summary: 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).
topic Cell Biology
Developmental biology
Gene Expression
In Situ Hybridization
Microscopy
Model Organisms
url http://www.sciencedirect.com/science/article/pii/S2666166721005189
work_keys_str_mv AT daisyjvinter liveandfixedimagingoftranslationsitesatsinglemrnaresolutioninthedrosophilaembryo
AT carolinehoppe liveandfixedimagingoftranslationsitesatsinglemrnaresolutioninthedrosophilaembryo
AT hilarylashe liveandfixedimagingoftranslationsitesatsinglemrnaresolutioninthedrosophilaembryo
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