Cellular reporter for inflammation.
Inflammatory response in cells are often measured through expression of a key transcription factor called NF-B. Currently, there are commercial green fluorescent protein (GFP) assays available for in vitro cell lines only. There are no in vivo systems that can report NF-B activation in hematopoiet...
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| Format: | Final Year Project (FYP) |
| Language: | English |
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2011
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| Online Access: | http://hdl.handle.net/10356/45263 |
| _version_ | 1824454142649696256 |
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| author | Yuhanis Abdullah Sunny. |
| author2 | Klaus Erik Karjalainen |
| author_facet | Klaus Erik Karjalainen Yuhanis Abdullah Sunny. |
| author_sort | Yuhanis Abdullah Sunny. |
| collection | NTU |
| description | Inflammatory response in cells are often measured through expression of a key transcription factor called NF-B. Currently, there are commercial green fluorescent protein (GFP) assays available for in vitro cell lines only. There are no in vivo systems that can report NF-B activation in hematopoietic stem cells. As such, this project is aimed at trying to develop such an in vivo assay, albeit at a preliminary stage. We have a retroviral reporter for NF-B in which we conducted experiments to test the workability of this reporter. Through a process called retroviral-mediated gene transfer, we transduced bone marrow cells with the reporter and tested for GFP expression in presence of pro-inflammatory stimuli. We had found that our reporter could clearly show GFP expression well when the transduced bone marrow cells were induced to undergo inflammation. However, upon differentiating the transduced bone marrow cells into macrophage and stimulating to induce inflammation, GFP expression was low which we suspect was due to a silencing of the reporter during differentiation. Further experiments need to be done in vivo to confirm whether the reporter is able to express GFP in presence of pro-inflammatory stimuli. |
| first_indexed | 2025-02-19T03:17:37Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/45263 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2025-02-19T03:17:37Z |
| publishDate | 2011 |
| record_format | dspace |
| spelling | ntu-10356/452632023-02-28T18:01:26Z Cellular reporter for inflammation. Yuhanis Abdullah Sunny. Klaus Erik Karjalainen School of Biological Sciences DRNTU::Science::Biological sciences::Microbiology::Immunology Inflammatory response in cells are often measured through expression of a key transcription factor called NF-B. Currently, there are commercial green fluorescent protein (GFP) assays available for in vitro cell lines only. There are no in vivo systems that can report NF-B activation in hematopoietic stem cells. As such, this project is aimed at trying to develop such an in vivo assay, albeit at a preliminary stage. We have a retroviral reporter for NF-B in which we conducted experiments to test the workability of this reporter. Through a process called retroviral-mediated gene transfer, we transduced bone marrow cells with the reporter and tested for GFP expression in presence of pro-inflammatory stimuli. We had found that our reporter could clearly show GFP expression well when the transduced bone marrow cells were induced to undergo inflammation. However, upon differentiating the transduced bone marrow cells into macrophage and stimulating to induce inflammation, GFP expression was low which we suspect was due to a silencing of the reporter during differentiation. Further experiments need to be done in vivo to confirm whether the reporter is able to express GFP in presence of pro-inflammatory stimuli. Bachelor of Science in Biological Sciences 2011-06-10T06:12:21Z 2011-06-10T06:12:21Z 2011 2011 Final Year Project (FYP) http://hdl.handle.net/10356/45263 en Nanyang Technological University 26 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences::Microbiology::Immunology Yuhanis Abdullah Sunny. Cellular reporter for inflammation. |
| title | Cellular reporter for inflammation. |
| title_full | Cellular reporter for inflammation. |
| title_fullStr | Cellular reporter for inflammation. |
| title_full_unstemmed | Cellular reporter for inflammation. |
| title_short | Cellular reporter for inflammation. |
| title_sort | cellular reporter for inflammation |
| topic | DRNTU::Science::Biological sciences::Microbiology::Immunology |
| url | http://hdl.handle.net/10356/45263 |
| work_keys_str_mv | AT yuhanisabdullahsunny cellularreporterforinflammation |