Gene therapy using viral vector against anxiety

• Main Author: Loo, Sze Jie
• Other Authors: Rupshi Mitra
• Format: Final Year Project
• Language: English
• Published: 2016
• Subjects: DRNTU::Science
• Online Access: http://hdl.handle.net/10356/67358
• Institution: Nanyang Technological University

Anxiety is an emotion that encompasses anxiousness and nervousness which can develop into neurological disorders caused by chronic stressors. Anxiety is mainly mediated by the amygdala that positively regulates the HPA axis, resulting in elevated levels of stress hormones in a person. A heightened HPA axis can further promote neuronal growth and increase spine density in the amygdala, forming a vicious cycle. One of the growth factors that modulate this development is Brain-Derived Neurotrophic Factor (BDNF). In this study, it was hypothesized that over-expression of BDNF in the BLA can lead to anxiety disorders. To prove this, the aim of the study was to clone BDNF gene into AAV vector, allowing co-transfection with pRC-AAV and phelper-AAV plasmids into HEK293T cells to generate viral particles. This project has been optimising BDNF gene cloning and viral packaging protocols to allow the continuation of BDNF gene research. Although unsuccessful in gene cloning, solutions were provided to rectify possible issues in the protocol. Future studies such as BDNF gene silencing and anxiety-related behaviour studies will be needed to support the hypothesis that BDNF gene is a major contributor in causing anxiety.