Generation and analysis of mouse embryonic stem cells with knockout of the <i>Mcph1</i> (microcephalin) gene
Chromatin is not randomly distributed within the nucleus, but organized in a three-dimensional structure that plays a critical role in genome functions. Сohesin and condensins are conserved multi-subunit protein complexes that participate in mammalian genome organization by extruding chromatin loops...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders
2024-09-01
|
| Series: | Вавиловский журнал генетики и селекции |
| Subjects: | |
| Online Access: | https://vavilov.elpub.ru/jour/article/view/4230 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832575051805753344 |
|---|---|
| author | A. M. Yunusova A. V. Smirnov T. A. Shnaider I. E. Pristyazhnuk S. Y. Korableva N. R. Battulin |
| author_facet | A. M. Yunusova A. V. Smirnov T. A. Shnaider I. E. Pristyazhnuk S. Y. Korableva N. R. Battulin |
| author_sort | A. M. Yunusova |
| collection | DOAJ |
| description | Chromatin is not randomly distributed within the nucleus, but organized in a three-dimensional structure that plays a critical role in genome functions. Сohesin and condensins are conserved multi-subunit protein complexes that participate in mammalian genome organization by extruding chromatin loops. The fine temporal regulation of these complexes is facilitated by a number of other proteins, one of which is microcephalin (Mcph1). Mcph1 prevents condensin II from associating with chromatin through interphase. Loss of Mcph1 induces chromosome hypercondensation; it is not clear to what extent this reorganization affects gene expression. In this study, we generated several mouse embryonic stem cell (mESC) lines with knockout of the Mcph1 gene and analyzed their gene expression profile. Gene Ontology analyses of differentially expressed genes (DEGs) after Mcph1 knockout revealed gene categories related to general metabolism and olfactory receptor function but not to cell cycle control previously described for Mcph1. We did not find a correlation between the DEGs and their frequency of lamina association. Thus, this evidence questions the hypothesis that Mcph1 knockout-mediated chromatin reorganization governs gene expression in mESCs. Among the negative effects of Mcph1 knockout, we observed numerous chromosomal aberrations, including micronucleus formation and chromosome fusion. This confirms the role of Mcph1 in maintaining genome integrity described previously. In our opinion, dysfunction of Mcph1 may be a kind of “Rosetta stone” for deciphering the function of condensin II in the interphase nucleus. Thus, the cell lines with knocked-out Mcph1 can be used to further study the influence of chromatin structural proteins on gene expression. |
| format | Article |
| id | doaj-art-e79b03123ed844f8a7f415e53718885c |
| institution | Kabale University |
| issn | 2500-3259 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders |
| record_format | Article |
| series | Вавиловский журнал генетики и селекции |
| spelling | doaj-art-e79b03123ed844f8a7f415e53718885c2025-02-01T09:58:13ZengSiberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and BreedersВавиловский журнал генетики и селекции2500-32592024-09-0128548749410.18699/vjgb-24-551483Generation and analysis of mouse embryonic stem cells with knockout of the <i>Mcph1</i> (microcephalin) geneA. M. Yunusova0A. V. Smirnov1T. A. Shnaider2I. E. Pristyazhnuk3S. Y. Korableva4N. R. Battulin5Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of SciencesInstitute of Cytology and Genetics of the Siberian Branch of the Russian Academy of SciencesInstitute of Cytology and Genetics of the Siberian Branch of the Russian Academy of SciencesInstitute of Cytology and Genetics of the Siberian Branch of the Russian Academy of SciencesNovosibirsk State UniversityInstitute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences; Novosibirsk State UniversityChromatin is not randomly distributed within the nucleus, but organized in a three-dimensional structure that plays a critical role in genome functions. Сohesin and condensins are conserved multi-subunit protein complexes that participate in mammalian genome organization by extruding chromatin loops. The fine temporal regulation of these complexes is facilitated by a number of other proteins, one of which is microcephalin (Mcph1). Mcph1 prevents condensin II from associating with chromatin through interphase. Loss of Mcph1 induces chromosome hypercondensation; it is not clear to what extent this reorganization affects gene expression. In this study, we generated several mouse embryonic stem cell (mESC) lines with knockout of the Mcph1 gene and analyzed their gene expression profile. Gene Ontology analyses of differentially expressed genes (DEGs) after Mcph1 knockout revealed gene categories related to general metabolism and olfactory receptor function but not to cell cycle control previously described for Mcph1. We did not find a correlation between the DEGs and their frequency of lamina association. Thus, this evidence questions the hypothesis that Mcph1 knockout-mediated chromatin reorganization governs gene expression in mESCs. Among the negative effects of Mcph1 knockout, we observed numerous chromosomal aberrations, including micronucleus formation and chromosome fusion. This confirms the role of Mcph1 in maintaining genome integrity described previously. In our opinion, dysfunction of Mcph1 may be a kind of “Rosetta stone” for deciphering the function of condensin II in the interphase nucleus. Thus, the cell lines with knocked-out Mcph1 can be used to further study the influence of chromatin structural proteins on gene expression.https://vavilov.elpub.ru/jour/article/view/4230mcph1/microcephalinchromosome condensationmescsgene expression analysis |
| spellingShingle | A. M. Yunusova A. V. Smirnov T. A. Shnaider I. E. Pristyazhnuk S. Y. Korableva N. R. Battulin Generation and analysis of mouse embryonic stem cells with knockout of the <i>Mcph1</i> (microcephalin) gene Вавиловский журнал генетики и селекции mcph1/microcephalin chromosome condensation mescs gene expression analysis |
| title | Generation and analysis of mouse embryonic stem cells with knockout of the <i>Mcph1</i> (microcephalin) gene |
| title_full | Generation and analysis of mouse embryonic stem cells with knockout of the <i>Mcph1</i> (microcephalin) gene |
| title_fullStr | Generation and analysis of mouse embryonic stem cells with knockout of the <i>Mcph1</i> (microcephalin) gene |
| title_full_unstemmed | Generation and analysis of mouse embryonic stem cells with knockout of the <i>Mcph1</i> (microcephalin) gene |
| title_short | Generation and analysis of mouse embryonic stem cells with knockout of the <i>Mcph1</i> (microcephalin) gene |
| title_sort | generation and analysis of mouse embryonic stem cells with knockout of the i mcph1 i microcephalin gene |
| topic | mcph1/microcephalin chromosome condensation mescs gene expression analysis |
| url | https://vavilov.elpub.ru/jour/article/view/4230 |
| work_keys_str_mv | AT amyunusova generationandanalysisofmouseembryonicstemcellswithknockoutoftheimcph1imicrocephalingene AT avsmirnov generationandanalysisofmouseembryonicstemcellswithknockoutoftheimcph1imicrocephalingene AT tashnaider generationandanalysisofmouseembryonicstemcellswithknockoutoftheimcph1imicrocephalingene AT iepristyazhnuk generationandanalysisofmouseembryonicstemcellswithknockoutoftheimcph1imicrocephalingene AT sykorableva generationandanalysisofmouseembryonicstemcellswithknockoutoftheimcph1imicrocephalingene AT nrbattulin generationandanalysisofmouseembryonicstemcellswithknockoutoftheimcph1imicrocephalingene |