Deciphering the Nature of <i>Trp</i>73 Isoforms in Mouse Embryonic Stem Cell Models: Generation of Isoform-Specific Deficient Cell Lines Using the CRISPR/Cas9 Gene Editing System
The p53 family has been widely studied for its role in various physiological and pathological processes. Imbalance of p53 family proteins may contribute to developmental abnormalities and pathologies in humans. This family exerts its functions through a profusion of isoforms that are generated by di...
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MDPI AG
2021-06-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/13/13/3182 |
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| author | Lorena López-Ferreras Nicole Martínez-García Laura Maeso-Alonso Marta Martín-López Ángela Díez-Matilla Javier Villoch-Fernandez Hugo Alonso-Olivares Margarita M. Marques Maria C. Marin |
| author_facet | Lorena López-Ferreras Nicole Martínez-García Laura Maeso-Alonso Marta Martín-López Ángela Díez-Matilla Javier Villoch-Fernandez Hugo Alonso-Olivares Margarita M. Marques Maria C. Marin |
| author_sort | Lorena López-Ferreras |
| collection | DOAJ |
| description | The p53 family has been widely studied for its role in various physiological and pathological processes. Imbalance of p53 family proteins may contribute to developmental abnormalities and pathologies in humans. This family exerts its functions through a profusion of isoforms that are generated by different promoter usage and alternative splicing in a cell type dependent manner. In particular, the <i>Trp73</i> gene gives rise to TA and DN-p73 isoforms that confer p73 a dual nature. The biological relevance of p73 does not only rely on its tumor suppression effects, but on its pivotal role in several developmental processes. Therefore, the generation of cellular models that allow the study of the individual isoforms in a physiological context is of great biomedical relevance. We generated specific TA and DN-p73-deficient mouse embryonic stem cell lines using the CRISPR/Cas9 gene editing system and validated them as physiological bona fide p73-isoform knockout models. Global gene expression analysis revealed isoform-specific alterations of distinctive transcriptional networks. Elimination of TA or DN-p73 is compatible with pluripotency but prompts naïve pluripotent stem cell transition into the primed state, compromising adequate lineage differentiation, thus suggesting that differential expression of p73 isoforms acts as a rheostat during early cell fate determination. |
| first_indexed | 2024-03-10T10:02:55Z |
| format | Article |
| id | doaj.art-b93c8abdb422413db2b8a61002babf90 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-10T10:02:55Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-b93c8abdb422413db2b8a61002babf902023-11-22T01:48:17ZengMDPI AGCancers2072-66942021-06-011313318210.3390/cancers13133182Deciphering the Nature of <i>Trp</i>73 Isoforms in Mouse Embryonic Stem Cell Models: Generation of Isoform-Specific Deficient Cell Lines Using the CRISPR/Cas9 Gene Editing SystemLorena López-Ferreras0Nicole Martínez-García1Laura Maeso-Alonso2Marta Martín-López3Ángela Díez-Matilla4Javier Villoch-Fernandez5Hugo Alonso-Olivares6Margarita M. Marques7Maria C. Marin8Instituto de Biomedicina (IBIOMED), Universidad de León, 24071 León, SpainInstituto de Biomedicina (IBIOMED), Universidad de León, 24071 León, SpainInstituto de Biomedicina (IBIOMED), Universidad de León, 24071 León, SpainInstituto de Biomedicina (IBIOMED), Universidad de León, 24071 León, SpainInstituto de Biomedicina (IBIOMED), Universidad de León, 24071 León, SpainInstituto de Biomedicina (IBIOMED), Universidad de León, 24071 León, SpainInstituto de Biomedicina (IBIOMED), Universidad de León, 24071 León, SpainDepartamento de Producción Animal, Universidad de León, 24071 León, SpainInstituto de Biomedicina (IBIOMED), Universidad de León, 24071 León, SpainThe p53 family has been widely studied for its role in various physiological and pathological processes. Imbalance of p53 family proteins may contribute to developmental abnormalities and pathologies in humans. This family exerts its functions through a profusion of isoforms that are generated by different promoter usage and alternative splicing in a cell type dependent manner. In particular, the <i>Trp73</i> gene gives rise to TA and DN-p73 isoforms that confer p73 a dual nature. The biological relevance of p73 does not only rely on its tumor suppression effects, but on its pivotal role in several developmental processes. Therefore, the generation of cellular models that allow the study of the individual isoforms in a physiological context is of great biomedical relevance. We generated specific TA and DN-p73-deficient mouse embryonic stem cell lines using the CRISPR/Cas9 gene editing system and validated them as physiological bona fide p73-isoform knockout models. Global gene expression analysis revealed isoform-specific alterations of distinctive transcriptional networks. Elimination of TA or DN-p73 is compatible with pluripotency but prompts naïve pluripotent stem cell transition into the primed state, compromising adequate lineage differentiation, thus suggesting that differential expression of p73 isoforms acts as a rheostat during early cell fate determination.https://www.mdpi.com/2072-6694/13/13/3182p73 isoformsTAp73DNp73mouse embryonic stem cells (mESC)gene editingpluripotency |
| spellingShingle | Lorena López-Ferreras Nicole Martínez-García Laura Maeso-Alonso Marta Martín-López Ángela Díez-Matilla Javier Villoch-Fernandez Hugo Alonso-Olivares Margarita M. Marques Maria C. Marin Deciphering the Nature of <i>Trp</i>73 Isoforms in Mouse Embryonic Stem Cell Models: Generation of Isoform-Specific Deficient Cell Lines Using the CRISPR/Cas9 Gene Editing System Cancers p73 isoforms TAp73 DNp73 mouse embryonic stem cells (mESC) gene editing pluripotency |
| title | Deciphering the Nature of <i>Trp</i>73 Isoforms in Mouse Embryonic Stem Cell Models: Generation of Isoform-Specific Deficient Cell Lines Using the CRISPR/Cas9 Gene Editing System |
| title_full | Deciphering the Nature of <i>Trp</i>73 Isoforms in Mouse Embryonic Stem Cell Models: Generation of Isoform-Specific Deficient Cell Lines Using the CRISPR/Cas9 Gene Editing System |
| title_fullStr | Deciphering the Nature of <i>Trp</i>73 Isoforms in Mouse Embryonic Stem Cell Models: Generation of Isoform-Specific Deficient Cell Lines Using the CRISPR/Cas9 Gene Editing System |
| title_full_unstemmed | Deciphering the Nature of <i>Trp</i>73 Isoforms in Mouse Embryonic Stem Cell Models: Generation of Isoform-Specific Deficient Cell Lines Using the CRISPR/Cas9 Gene Editing System |
| title_short | Deciphering the Nature of <i>Trp</i>73 Isoforms in Mouse Embryonic Stem Cell Models: Generation of Isoform-Specific Deficient Cell Lines Using the CRISPR/Cas9 Gene Editing System |
| title_sort | deciphering the nature of i trp i 73 isoforms in mouse embryonic stem cell models generation of isoform specific deficient cell lines using the crispr cas9 gene editing system |
| topic | p73 isoforms TAp73 DNp73 mouse embryonic stem cells (mESC) gene editing pluripotency |
| url | https://www.mdpi.com/2072-6694/13/13/3182 |
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