A method for detecting single mRNA molecules in Arabidopsis thaliana

BACKGROUND: Despite advances in other model organisms, there are currently no techniques to explore cell-to-cell variation and sub-cellular localization of RNA molecules at the single-cell level in plants. RESULTS: Here we describe a method for imaging individual mRNA molecules in Arabidopsis thalia...

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Detalles Bibliográficos
Autores principales: Duncan, Susan, Olsson, Tjelvar S. G., Hartley, Matthew, Dean, Caroline, Rosa, Stefanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Methodology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5192599/
https://www.ncbi.nlm.nih.gov/pubmed/28035231
http://dx.doi.org/10.1186/s13007-016-0114-x
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author Duncan, Susan
Olsson, Tjelvar S. G.
Hartley, Matthew
Dean, Caroline
Rosa, Stefanie
author_facet Duncan, Susan
Olsson, Tjelvar S. G.
Hartley, Matthew
Dean, Caroline
Rosa, Stefanie
author_sort Duncan, Susan
collection PubMed
description BACKGROUND: Despite advances in other model organisms, there are currently no techniques to explore cell-to-cell variation and sub-cellular localization of RNA molecules at the single-cell level in plants. RESULTS: Here we describe a method for imaging individual mRNA molecules in Arabidopsis thaliana root cells using multiple singly labeled oligonucleotide probes. We demonstrate detection of both mRNA and nascent transcripts of the housekeeping gene Protein Phosphatase 2A. Our image analysis pipeline also enables quantification of mRNAs that reveals the frequency distribution of transcripts per cell underlying the population mean. CONCLUSION: This method allows single molecule RNA in situ to be exploited as a powerful tool for studying gene regulation in plants. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13007-016-0114-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-51925992016-12-29 A method for detecting single mRNA molecules in Arabidopsis thaliana Duncan, Susan Olsson, Tjelvar S. G. Hartley, Matthew Dean, Caroline Rosa, Stefanie Plant Methods Methodology BACKGROUND: Despite advances in other model organisms, there are currently no techniques to explore cell-to-cell variation and sub-cellular localization of RNA molecules at the single-cell level in plants. RESULTS: Here we describe a method for imaging individual mRNA molecules in Arabidopsis thaliana root cells using multiple singly labeled oligonucleotide probes. We demonstrate detection of both mRNA and nascent transcripts of the housekeeping gene Protein Phosphatase 2A. Our image analysis pipeline also enables quantification of mRNAs that reveals the frequency distribution of transcripts per cell underlying the population mean. CONCLUSION: This method allows single molecule RNA in situ to be exploited as a powerful tool for studying gene regulation in plants. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13007-016-0114-x) contains supplementary material, which is available to authorized users. BioMed Central 2016-08-05 /pmc/articles/PMC5192599/ /pubmed/28035231 http://dx.doi.org/10.1186/s13007-016-0114-x Text en © Duncan et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Duncan, Susan
Olsson, Tjelvar S. G.
Hartley, Matthew
Dean, Caroline
Rosa, Stefanie
A method for detecting single mRNA molecules in Arabidopsis thaliana
title A method for detecting single mRNA molecules in Arabidopsis thaliana
title_full A method for detecting single mRNA molecules in Arabidopsis thaliana
title_fullStr A method for detecting single mRNA molecules in Arabidopsis thaliana
title_full_unstemmed A method for detecting single mRNA molecules in Arabidopsis thaliana
title_short A method for detecting single mRNA molecules in Arabidopsis thaliana
title_sort method for detecting single mrna molecules in arabidopsis thaliana
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5192599/
https://www.ncbi.nlm.nih.gov/pubmed/28035231
http://dx.doi.org/10.1186/s13007-016-0114-x
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