Huntington’s disease modeling on HEK293 cell line
Huntington’s disease is a hereditary neurodegenerative disorder caused by CAG trinucleotide repeat expansion in the first exon of HTT gene. The mutant HTT protein has an elongated polyglutamine tract and forms aggregates in the nuclei and cytoplasm of the striatal neurons. The pathological processes...
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| Format: | Article |
| Language: | English |
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Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders
2017-12-01
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| Series: | Вавиловский журнал генетики и селекции |
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| Online Access: | https://vavilov.elpub.ru/jour/article/view/1236 |
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| author | D. V. Sharipova T. B. Malankhanova A. A. Malakhova |
| author_facet | D. V. Sharipova T. B. Malankhanova A. A. Malakhova |
| author_sort | D. V. Sharipova |
| collection | DOAJ |
| description | Huntington’s disease is a hereditary neurodegenerative disorder caused by CAG trinucleotide repeat expansion in the first exon of HTT gene. The mutant HTT protein has an elongated polyglutamine tract and forms aggregates in the nuclei and cytoplasm of the striatal neurons. The pathological processes occurring in the medium spiny neurons of Huntington’s disease patients lead to neurodegeneration and consequently to the death. The molecular mechanisms of the pathology development are difficult to study due to the limited material availability and late onset of the manifestation. Therefore, one of the important tasks is generation of an in vitro model system of Huntington’s disease based on human cell cultures. The new genome editing approaches, such as CRISPR/Cas9, allow us to generate isogenic cell lines that can be useful for drug screening and studying mechanisms of molecular and cellular events triggered by certain mutation on an equal genetic background. Here, we investigated the viability and proliferative rate of several mutant HEK293 cell clones with mutations in the first exon of HTT gene. The mutant clones were obtained earlier using CRISPR/Cas9 genome editing technology. We showed that mutant cells partially reproduce the pathological phenotype, that is, they have reduced proliferation activity, an increased level of apoptosis and high sensitivity to treatment with 5μM MG132 proteasome inhibitor compared to the original HEK293 Phoenix cell line. Our results indicate that the mutation in the first exon of HTT gene affects not only neurons, but also other types of cells, and HEK293 cell clones bearing the mutation can serve as in vitro model for studying some mechanisms of HTT functioning. |
| first_indexed | 2024-03-07T16:06:54Z |
| format | Article |
| id | doaj.art-31c4924c4a68462f9c31cc65c2d7eca5 |
| institution | Directory Open Access Journal |
| issn | 2500-3259 |
| language | English |
| last_indexed | 2024-04-24T11:08:13Z |
| publishDate | 2017-12-01 |
| 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-31c4924c4a68462f9c31cc65c2d7eca52024-04-11T15:30:58ZengSiberian 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 селекции2500-32592017-12-0121785686110.18699/VJ17.306694Huntington’s disease modeling on HEK293 cell lineD. V. Sharipova0T. B. Malankhanova1A. A. Malakhova2Novosibirsk State University; Institute of Cytology and Genetics SB RAS; Meshalkin National Medical Research Center of the Ministry of Healthcare of the Russian Federation.Novosibirsk State University; Institute of Cytology and Genetics SB RAS; Meshalkin National Medical Research Center of the Ministry of Healthcare of the Russian Federation; Institute of Chemical Biology and Fundamental Medicine SB RAS.Novosibirsk State University; Institute of Cytology and Genetics SB RAS; Meshalkin National Medical Research Center of the Ministry of Healthcare of the Russian Federation; Institute of Chemical Biology and Fundamental Medicine SB RAS.Huntington’s disease is a hereditary neurodegenerative disorder caused by CAG trinucleotide repeat expansion in the first exon of HTT gene. The mutant HTT protein has an elongated polyglutamine tract and forms aggregates in the nuclei and cytoplasm of the striatal neurons. The pathological processes occurring in the medium spiny neurons of Huntington’s disease patients lead to neurodegeneration and consequently to the death. The molecular mechanisms of the pathology development are difficult to study due to the limited material availability and late onset of the manifestation. Therefore, one of the important tasks is generation of an in vitro model system of Huntington’s disease based on human cell cultures. The new genome editing approaches, such as CRISPR/Cas9, allow us to generate isogenic cell lines that can be useful for drug screening and studying mechanisms of molecular and cellular events triggered by certain mutation on an equal genetic background. Here, we investigated the viability and proliferative rate of several mutant HEK293 cell clones with mutations in the first exon of HTT gene. The mutant clones were obtained earlier using CRISPR/Cas9 genome editing technology. We showed that mutant cells partially reproduce the pathological phenotype, that is, they have reduced proliferation activity, an increased level of apoptosis and high sensitivity to treatment with 5μM MG132 proteasome inhibitor compared to the original HEK293 Phoenix cell line. Our results indicate that the mutation in the first exon of HTT gene affects not only neurons, but also other types of cells, and HEK293 cell clones bearing the mutation can serve as in vitro model for studying some mechanisms of HTT functioning.https://vavilov.elpub.ru/jour/article/view/1236huntington’s diseasecell modelsgenome editing |
| spellingShingle | D. V. Sharipova T. B. Malankhanova A. A. Malakhova Huntington’s disease modeling on HEK293 cell line Вавиловский журнал генетики и селекции huntington’s disease cell models genome editing |
| title | Huntington’s disease modeling on HEK293 cell line |
| title_full | Huntington’s disease modeling on HEK293 cell line |
| title_fullStr | Huntington’s disease modeling on HEK293 cell line |
| title_full_unstemmed | Huntington’s disease modeling on HEK293 cell line |
| title_short | Huntington’s disease modeling on HEK293 cell line |
| title_sort | huntington s disease modeling on hek293 cell line |
| topic | huntington’s disease cell models genome editing |
| url | https://vavilov.elpub.ru/jour/article/view/1236 |
| work_keys_str_mv | AT dvsharipova huntingtonsdiseasemodelingonhek293cellline AT tbmalankhanova huntingtonsdiseasemodelingonhek293cellline AT aamalakhova huntingtonsdiseasemodelingonhek293cellline |