Validation of candidate genes involved in X chromosome inactivation
One of the two female X chromosomes is silenced in mammals to compensate for X-linked gene dosage. For over half a century, this process, known as X chromosome inactivation (XCI), has been studied intensively and Xist was found to have a central role in it. Although Xist RNA is critical for establis...
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| Formato: | Final Year Project (FYP) |
| Idioma: | English |
| Publicado em: |
2015
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| Acesso em linha: | http://hdl.handle.net/10356/63613 |
| _version_ | 1826127814113361920 |
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| author | Lim, Zhong Ri |
| author2 | Zhang Li Feng |
| author_facet | Zhang Li Feng Lim, Zhong Ri |
| author_sort | Lim, Zhong Ri |
| collection | NTU |
| description | One of the two female X chromosomes is silenced in mammals to compensate for X-linked gene dosage. For over half a century, this process, known as X chromosome inactivation (XCI), has been studied intensively and Xist was found to have a central role in it. Although Xist RNA is critical for establishing XCI during early embryonic development, it plays a dispensable role in maintaining XCI in soma. In order to discover unknown factors involved in XCI, our lab carried out a genome-wide shRNA library screen. In this study, three candidate genes, M3, Kdm5c and MUL1, were validated individually. Using CRISPR/Cas9 system, gene knockout was carried out on AV3.1.12 cell line, which is a male mouse ES cell line carrying an inducible Xist transgene along its single male X chromosome. Inducible Xist expression causes severe cell death of AV3.1.12 cells due to ectopic XCI. As a result, knocking out a gene involved in XCI would rescue the cells. While knocking out Kdm5c and MUL1 did not rescue the cells, knocking out M3 showed a significant rescuing effect on AV3.1.12 cells. Consequently, the role of M3 in XCI is being further confirmed and investigated. |
| first_indexed | 2024-10-01T07:14:37Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/63613 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:14:37Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | ntu-10356/636132023-02-28T18:01:13Z Validation of candidate genes involved in X chromosome inactivation Lim, Zhong Ri Zhang Li Feng School of Biological Sciences DRNTU::Science::Biological sciences One of the two female X chromosomes is silenced in mammals to compensate for X-linked gene dosage. For over half a century, this process, known as X chromosome inactivation (XCI), has been studied intensively and Xist was found to have a central role in it. Although Xist RNA is critical for establishing XCI during early embryonic development, it plays a dispensable role in maintaining XCI in soma. In order to discover unknown factors involved in XCI, our lab carried out a genome-wide shRNA library screen. In this study, three candidate genes, M3, Kdm5c and MUL1, were validated individually. Using CRISPR/Cas9 system, gene knockout was carried out on AV3.1.12 cell line, which is a male mouse ES cell line carrying an inducible Xist transgene along its single male X chromosome. Inducible Xist expression causes severe cell death of AV3.1.12 cells due to ectopic XCI. As a result, knocking out a gene involved in XCI would rescue the cells. While knocking out Kdm5c and MUL1 did not rescue the cells, knocking out M3 showed a significant rescuing effect on AV3.1.12 cells. Consequently, the role of M3 in XCI is being further confirmed and investigated. Bachelor of Science in Biological Sciences 2015-05-18T01:19:12Z 2015-05-18T01:19:12Z 2015 2015 Final Year Project (FYP) http://hdl.handle.net/10356/63613 en 35 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences Lim, Zhong Ri Validation of candidate genes involved in X chromosome inactivation |
| title | Validation of candidate genes involved in X chromosome inactivation |
| title_full | Validation of candidate genes involved in X chromosome inactivation |
| title_fullStr | Validation of candidate genes involved in X chromosome inactivation |
| title_full_unstemmed | Validation of candidate genes involved in X chromosome inactivation |
| title_short | Validation of candidate genes involved in X chromosome inactivation |
| title_sort | validation of candidate genes involved in x chromosome inactivation |
| topic | DRNTU::Science::Biological sciences |
| url | http://hdl.handle.net/10356/63613 |
| work_keys_str_mv | AT limzhongri validationofcandidategenesinvolvedinxchromosomeinactivation |