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Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR
mRNA decay is an important process in post-transcriptional regulation; in addition, it plays a crucial role in plant development and response to stress. The development of new tools to quantify mRNA decay intermediates is thus important to better characterize the dynamic of mRNA decay in various con...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9571659/ https://www.ncbi.nlm.nih.gov/pubmed/36235485 http://dx.doi.org/10.3390/plants11192616 |
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author | Boubegtitene, Alexandre Merret, Rémy |
author_facet | Boubegtitene, Alexandre Merret, Rémy |
author_sort | Boubegtitene, Alexandre |
collection | PubMed |
description | mRNA decay is an important process in post-transcriptional regulation; in addition, it plays a crucial role in plant development and response to stress. The development of new tools to quantify mRNA decay intermediates is thus important to better characterize the dynamic of mRNA decay in various conditions. Here, we applied droplet digital PCR (ddPCR), a recent and precise PCR technology, to determine mRNA half-life in Arabidopsis seedlings. We demonstrated that ddPCR can correctly assess mRNA half-life from a wide variety of transcripts in a reproducible manner. We also demonstrated that thanks to multiplexing mRNA, the half-life of multiple transcripts can be followed in the same reaction. As ddPCR allows precise quantification, we proposed that this approach is highly suitable when a low amount of RNA is available; for the detection of many targets or for the analysis of lowly expressed transcripts. |
format | Online Article Text |
id | pubmed-9571659 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-95716592022-10-17 Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR Boubegtitene, Alexandre Merret, Rémy Plants (Basel) Technical Note mRNA decay is an important process in post-transcriptional regulation; in addition, it plays a crucial role in plant development and response to stress. The development of new tools to quantify mRNA decay intermediates is thus important to better characterize the dynamic of mRNA decay in various conditions. Here, we applied droplet digital PCR (ddPCR), a recent and precise PCR technology, to determine mRNA half-life in Arabidopsis seedlings. We demonstrated that ddPCR can correctly assess mRNA half-life from a wide variety of transcripts in a reproducible manner. We also demonstrated that thanks to multiplexing mRNA, the half-life of multiple transcripts can be followed in the same reaction. As ddPCR allows precise quantification, we proposed that this approach is highly suitable when a low amount of RNA is available; for the detection of many targets or for the analysis of lowly expressed transcripts. MDPI 2022-10-05 /pmc/articles/PMC9571659/ /pubmed/36235485 http://dx.doi.org/10.3390/plants11192616 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Technical Note Boubegtitene, Alexandre Merret, Rémy Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR |
title | Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR |
title_full | Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR |
title_fullStr | Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR |
title_full_unstemmed | Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR |
title_short | Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR |
title_sort | monitoring mrna half-life in arabidopsis using droplet digital pcr |
topic | Technical Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9571659/ https://www.ncbi.nlm.nih.gov/pubmed/36235485 http://dx.doi.org/10.3390/plants11192616 |
work_keys_str_mv | AT boubegtitenealexandre monitoringmrnahalflifeinarabidopsisusingdropletdigitalpcr AT merretremy monitoringmrnahalflifeinarabidopsisusingdropletdigitalpcr |