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CRISPR/Cas9 genome editing system confirms centriolin’s role in cytokinesis

OBJECTIVE: In addition to its function as the microtubule organizing center of the cell, the centrosome has functions in many other cellular processes including primary cilia formation, DNA damage checkpoints, and cell cycle progression. But the role of individual components of the centrosome in the...

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Autores principales: Seronick, Eric, Son, Jae, Michael, Cameron, Fogg, Hannah, Gromley, Zeynep, Gromley, Adam
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8751070/
https://www.ncbi.nlm.nih.gov/pubmed/35012607
http://dx.doi.org/10.1186/s13104-021-05898-w
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author Seronick, Eric
Son, Jae
Michael, Cameron
Fogg, Hannah
Gromley, Zeynep
Gromley, Adam
author_facet Seronick, Eric
Son, Jae
Michael, Cameron
Fogg, Hannah
Gromley, Zeynep
Gromley, Adam
author_sort Seronick, Eric
collection PubMed
description OBJECTIVE: In addition to its function as the microtubule organizing center of the cell, the centrosome has functions in many other cellular processes including primary cilia formation, DNA damage checkpoints, and cell cycle progression. But the role of individual components of the centrosome in these processes remains unclear. Previous studies used siRNA (small interfering RNA) to “knock down” protein levels of the centrosome component centriolin, resulting in failed cytokinesis. Since this approach was transient, only targeting centriolin at the mRNA level, we sought to confirm these findings by permanently disrupting the gene encoding centriolin using the CRISPR/Cas9 system of genome editing. RESULTS: This study provides evidence that the CRISPR/Cas9 system is capable of effectively reducing centriolin protein levels in the cell. Furthermore, this disruption leads to a failure of cytokinesis that is reminiscent of the phenotype previously reported for the siRNA-mediated disruption of centriolin. Furthermore, no additional defects in cell division were observed, consistent with results seen with previous siRNA studies. We conclude that the CRISPR/Cas9 system is an effective means of permanently removing the cellular pools of centriolin and that the disruption of centriolin at both the mRNA level and genomic level lead to similar cell division defects. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13104-021-05898-w.
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spelling pubmed-87510702022-01-11 CRISPR/Cas9 genome editing system confirms centriolin’s role in cytokinesis Seronick, Eric Son, Jae Michael, Cameron Fogg, Hannah Gromley, Zeynep Gromley, Adam BMC Res Notes Research Note OBJECTIVE: In addition to its function as the microtubule organizing center of the cell, the centrosome has functions in many other cellular processes including primary cilia formation, DNA damage checkpoints, and cell cycle progression. But the role of individual components of the centrosome in these processes remains unclear. Previous studies used siRNA (small interfering RNA) to “knock down” protein levels of the centrosome component centriolin, resulting in failed cytokinesis. Since this approach was transient, only targeting centriolin at the mRNA level, we sought to confirm these findings by permanently disrupting the gene encoding centriolin using the CRISPR/Cas9 system of genome editing. RESULTS: This study provides evidence that the CRISPR/Cas9 system is capable of effectively reducing centriolin protein levels in the cell. Furthermore, this disruption leads to a failure of cytokinesis that is reminiscent of the phenotype previously reported for the siRNA-mediated disruption of centriolin. Furthermore, no additional defects in cell division were observed, consistent with results seen with previous siRNA studies. We conclude that the CRISPR/Cas9 system is an effective means of permanently removing the cellular pools of centriolin and that the disruption of centriolin at both the mRNA level and genomic level lead to similar cell division defects. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13104-021-05898-w. BioMed Central 2022-01-10 /pmc/articles/PMC8751070/ /pubmed/35012607 http://dx.doi.org/10.1186/s13104-021-05898-w Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Note
Seronick, Eric
Son, Jae
Michael, Cameron
Fogg, Hannah
Gromley, Zeynep
Gromley, Adam
CRISPR/Cas9 genome editing system confirms centriolin’s role in cytokinesis
title CRISPR/Cas9 genome editing system confirms centriolin’s role in cytokinesis
title_full CRISPR/Cas9 genome editing system confirms centriolin’s role in cytokinesis
title_fullStr CRISPR/Cas9 genome editing system confirms centriolin’s role in cytokinesis
title_full_unstemmed CRISPR/Cas9 genome editing system confirms centriolin’s role in cytokinesis
title_short CRISPR/Cas9 genome editing system confirms centriolin’s role in cytokinesis
title_sort crispr/cas9 genome editing system confirms centriolin’s role in cytokinesis
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8751070/
https://www.ncbi.nlm.nih.gov/pubmed/35012607
http://dx.doi.org/10.1186/s13104-021-05898-w
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