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MUC4 is not expressed in cell lines used for live cell imaging

Background: The ability to visualise specific mammalian gene loci in living cells is important for understanding the dynamic processes linked to transcription. However, some of the tools used to target mammalian genes for live cell imaging, such as dCas9, have been reported to themselves impede proc...

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Autores principales: Athmane, Naouel, Williamson, Iain, Boyle, Shelagh, Biddie, Simon C., Bickmore, Wendy A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: F1000 Research Limited 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8567686/
https://www.ncbi.nlm.nih.gov/pubmed/34796278
http://dx.doi.org/10.12688/wellcomeopenres.17229.2
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author Athmane, Naouel
Williamson, Iain
Boyle, Shelagh
Biddie, Simon C.
Bickmore, Wendy A.
author_facet Athmane, Naouel
Williamson, Iain
Boyle, Shelagh
Biddie, Simon C.
Bickmore, Wendy A.
author_sort Athmane, Naouel
collection PubMed
description Background: The ability to visualise specific mammalian gene loci in living cells is important for understanding the dynamic processes linked to transcription. However, some of the tools used to target mammalian genes for live cell imaging, such as dCas9, have been reported to themselves impede processes linked to transcription. The MUC4 gene is a popular target for live cell imaging studies due to the repetitive nature of sequences within some exons of this gene. Methods: We set out to compare the impact of dCas9 and TALE-based imaging tools on MUC4 expression, including in human cell lines previously reported as expressing MUC4. Results: We were unable to detect MUC4 mRNA in these cell lines. Moreover, analysis of publicly available data for histone modifications associated with transcription, and data for transcription itself, indicate that neither MUC4, nor any of the mucin gene family are significantly expressed in the cell lines where dCas9 targeting has been reported to repress MUC4 and MUC1 expression, or in the cell lines where dCas13 has been used to report MUC4 RNA detection in live cells. Conclusions: Methods for visualising specific gene loci and gene transcripts in live human cells are very challenging. Our data suggest that care should be given to the choice of the most appropriate cell lines for these analyses and that orthogonal methods of assaying gene expression be carefully compared.
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spelling pubmed-85676862021-11-17 MUC4 is not expressed in cell lines used for live cell imaging Athmane, Naouel Williamson, Iain Boyle, Shelagh Biddie, Simon C. Bickmore, Wendy A. Wellcome Open Res Research Article Background: The ability to visualise specific mammalian gene loci in living cells is important for understanding the dynamic processes linked to transcription. However, some of the tools used to target mammalian genes for live cell imaging, such as dCas9, have been reported to themselves impede processes linked to transcription. The MUC4 gene is a popular target for live cell imaging studies due to the repetitive nature of sequences within some exons of this gene. Methods: We set out to compare the impact of dCas9 and TALE-based imaging tools on MUC4 expression, including in human cell lines previously reported as expressing MUC4. Results: We were unable to detect MUC4 mRNA in these cell lines. Moreover, analysis of publicly available data for histone modifications associated with transcription, and data for transcription itself, indicate that neither MUC4, nor any of the mucin gene family are significantly expressed in the cell lines where dCas9 targeting has been reported to repress MUC4 and MUC1 expression, or in the cell lines where dCas13 has been used to report MUC4 RNA detection in live cells. Conclusions: Methods for visualising specific gene loci and gene transcripts in live human cells are very challenging. Our data suggest that care should be given to the choice of the most appropriate cell lines for these analyses and that orthogonal methods of assaying gene expression be carefully compared. F1000 Research Limited 2021-11-19 /pmc/articles/PMC8567686/ /pubmed/34796278 http://dx.doi.org/10.12688/wellcomeopenres.17229.2 Text en Copyright: © 2021 Athmane N et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Athmane, Naouel
Williamson, Iain
Boyle, Shelagh
Biddie, Simon C.
Bickmore, Wendy A.
MUC4 is not expressed in cell lines used for live cell imaging
title MUC4 is not expressed in cell lines used for live cell imaging
title_full MUC4 is not expressed in cell lines used for live cell imaging
title_fullStr MUC4 is not expressed in cell lines used for live cell imaging
title_full_unstemmed MUC4 is not expressed in cell lines used for live cell imaging
title_short MUC4 is not expressed in cell lines used for live cell imaging
title_sort muc4 is not expressed in cell lines used for live cell imaging
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8567686/
https://www.ncbi.nlm.nih.gov/pubmed/34796278
http://dx.doi.org/10.12688/wellcomeopenres.17229.2
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