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http://hdl.handle.net/11375/25320
Title: | Mapping the Protein Interactome of ASD-associated Genes by using BioID2 |
Authors: | Cheng, Anran |
Advisor: | Doble, Bradley Singh, Karun |
Department: | Biochemistry and Biomedical Sciences |
Publication Date: | 2019 |
Abstract: | Autism spectrum disorder (ASD) is an array of neurodevelopmental disorders that vary in the severity of symptoms presented by affected individuals. In Canada, ASD is estimated to affect 1 in 66 children1. Though severity can vary, the clinical diagnostic criteria define deficits in social interaction, presence of repetitive behaviors and restricted interests, as characteristic manifestations2. The disorder not only reduces the quality of life for those affected, but also places an immense emotional and financial burden on their families and an economic burden on society. ASD is a heterogeneous disorder, with hundreds of implicated loci and genes. However, only a small number of implicated genes have well-defined neurobiological functions. Defining the pathogenesis and etiology of ASD in genetic and molecular terms is essential for expanding our understanding of the disorder, uncovering protein biomarkers for diagnosis, and identifying targets for therapeutic development. My project used a chemico-genetic proximity labeling approach, termed BioID2, to map the protein interactome of ASD-associated gene products. I employed the genome editing CRISPR-Cas9 technique to engineer mouse embryonic stem cells with a BioID2 expression cassette linked to an ASD-associated gene of interest. A mouse model will be generated from the edited mouse embryonic stem cells. This approach will permit the tracking of protein-protein interactions on an endogenous level, which provides a more accurate representation of the interacting partners. I also initiated a second approach in which lentiviral constructs harboring BioID2-tagged ASD-associated genes of interest will be used to overexpress resultant BioID2 fusion proteins in primary cortical neurons. This method will allow us to propose protein networks for our genes of interest and to dissect the molecular function of these genes. Studying protein-protein interactions of candidate ASD gene products is critical for understanding the molecular etiology of ASD. The elucidation of ASD-associated protein networks will be indispensable to the development of novel therapeutic agents, by informing target selection. This project will expand our current understanding of ASD, identify novel protein markers, and contribute to development of therapies. It can also serve as proof-of-concept that can be adapted for various other neurological disorders. |
URI: | http://hdl.handle.net/11375/25320 |
Appears in Collections: | Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
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Cheng_Anran_Thesis2019June_MSc.pdf | 2.07 MB | Adobe PDF | View/Open |
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