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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| Format: | Theses and Dissertations |
| Language: | English |
| Published: |
2019
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/87335 http://hdl.handle.net/10220/48048 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | 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. |
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