An inducible and reversible genetics platform for novel drug target discovery
Understanding gene function provides insights into biological processes and disease pathologies. Such knowledge can be applied to benefit human health and medicine. With this in mind, I have established a genetic platform in mouse to study gene function. The platform consists of two transgenic mo...
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| Other Authors: | |
| Format: | Thesis |
| Language: | English |
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2019
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| Online Access: | https://hdl.handle.net/10356/87335 http://hdl.handle.net/10220/48048 |
| _version_ | 1811685001998106624 |
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| author | Leong, Mei Ling |
| author2 | Klaus Erik Karjalainen |
| author_facet | Klaus Erik Karjalainen Leong, Mei Ling |
| author_sort | Leong, Mei Ling |
| collection | NTU |
| description | Understanding gene function provides insights into biological processes and disease
pathologies. Such knowledge can be applied to benefit human health and medicine.
With this in mind, I have established a genetic platform in mouse to study gene
function. The platform consists of two transgenic mouse strains that were generated inhouse,
through the use of CRISPR/Cas9 gene editing technology: (1) a strain that
ubiquitously expresses the reverse tetracycline trans-silencer (rTetR-KRAB) in all
tissues and (2) a strain where the tetracycline response element (TRE) has been inserted
in close proximity to the target gene’s promoter. With appropriate double transgenic
mouse strains, rTetR-KRAB binds to TRE in the presence of doxycycline and represses
the target gene. Repression is dose-dependent and reversible. To demonstrate the
feasibility of the platform, I have chosen to target Tcf7l2 which is a transcription factor
that acts downstream of Wnt signalling. The Wnt signalling pathway has been
implicated in colorectal cancer, but there are contradictory views on whether Tcf7l2 acts
as an oncogene or tumour suppressor gene in this context. Preliminary data obtained
using small intestine organoids derived from double transgenic mouse, show the
importance of Tcf7l2 for intestinal homeostasis and ApcMin-associated adenoma
maintenance. To expound this, I will utilise the Tcf7l2 transgenic strain to investigate if
physiological rebalance of this gene has therapeutic effects in colon cancer. This
platform can also be generally useful to study the physiological or pathological role of
other target genes. |
| first_indexed | 2024-10-01T04:37:35Z |
| format | Thesis |
| id | ntu-10356/87335 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:37:35Z |
| publishDate | 2019 |
| record_format | dspace |
| spelling | ntu-10356/873352023-02-28T18:38:03Z An inducible and reversible genetics platform for novel drug target discovery Leong, Mei Ling Klaus Erik Karjalainen School of Biological Sciences DRNTU::Science::Biological sciences Understanding gene function provides insights into biological processes and disease pathologies. Such knowledge can be applied to benefit human health and medicine. With this in mind, I have established a genetic platform in mouse to study gene function. The platform consists of two transgenic mouse strains that were generated inhouse, through the use of CRISPR/Cas9 gene editing technology: (1) a strain that ubiquitously expresses the reverse tetracycline trans-silencer (rTetR-KRAB) in all tissues and (2) a strain where the tetracycline response element (TRE) has been inserted in close proximity to the target gene’s promoter. With appropriate double transgenic mouse strains, rTetR-KRAB binds to TRE in the presence of doxycycline and represses the target gene. Repression is dose-dependent and reversible. To demonstrate the feasibility of the platform, I have chosen to target Tcf7l2 which is a transcription factor that acts downstream of Wnt signalling. The Wnt signalling pathway has been implicated in colorectal cancer, but there are contradictory views on whether Tcf7l2 acts as an oncogene or tumour suppressor gene in this context. Preliminary data obtained using small intestine organoids derived from double transgenic mouse, show the importance of Tcf7l2 for intestinal homeostasis and ApcMin-associated adenoma maintenance. To expound this, I will utilise the Tcf7l2 transgenic strain to investigate if physiological rebalance of this gene has therapeutic effects in colon cancer. This platform can also be generally useful to study the physiological or pathological role of other target genes. Doctor of Philosophy 2019-04-17T12:31:37Z 2019-12-06T16:39:44Z 2019-04-17T12:31:37Z 2019-12-06T16:39:44Z 2019 Thesis Leong, M. L. (2019). An inducible and reversible genetics platform for novel drug target discovery. Doctoral thesis, Nanyang Technological University, Singapore. https://hdl.handle.net/10356/87335 http://hdl.handle.net/10220/48048 10.32657/10220/48048 en 219 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences Leong, Mei Ling An inducible and reversible genetics platform for novel drug target discovery |
| title | An inducible and reversible genetics platform for novel drug target discovery |
| title_full | An inducible and reversible genetics platform for novel drug target discovery |
| title_fullStr | An inducible and reversible genetics platform for novel drug target discovery |
| title_full_unstemmed | An inducible and reversible genetics platform for novel drug target discovery |
| title_short | An inducible and reversible genetics platform for novel drug target discovery |
| title_sort | inducible and reversible genetics platform for novel drug target discovery |
| topic | DRNTU::Science::Biological sciences |
| url | https://hdl.handle.net/10356/87335 http://hdl.handle.net/10220/48048 |
| work_keys_str_mv | AT leongmeiling aninducibleandreversiblegeneticsplatformfornoveldrugtargetdiscovery AT leongmeiling inducibleandreversiblegeneticsplatformfornoveldrugtargetdiscovery |