Macquarie University
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Investigating amyotrophic lateral sclerosis candidate genes

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posted on 2022-03-29, 03:43 authored by Sandrine Kim Kiow Chan Moi Fat
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of motor neurons.Approximately 10% of cases are classiffed as familial and 90% sporadic, although a proportion of sporadic cases have an unrecognised family history. This thesis aimed o identify novel ALS genes from familial and sporadic cases. Whole-exome and whole genome sequencing analysis identified 22 candidate gene variants in an ALS family,which were investigated using an in silico analysis pipeline including protein predictions,conservation, genic tolerance, tissue expression and gene function. Three top priority variants, in CYB5R3, DCAF7, and SAV1, were assessed using in vitro toxicity and localisation assays, with one candidate, CYB5R3, displaying a differential cytoplasmic protein expression pattern. Whole-genome data from 635 sporadic ALS patients were interrogated for ALS candidate genes (n = 21), and ten novel variants were identified, including a novel CYB5R3 variant, which rated strongly from in silico tools. Lastly, this project sought to develop a CRISPR-Cas9 protocol to generate ALS cell models for future use in the in vitro analysis pipeline. Overall, a combination of genetic, in silico and in vitro pipelines were used to identify candidate genes in an ALS kindred and apparently sporadic patients. Novel ALS genes will further our understanding of disease biology and contribute to development of diagnostics and treatments.


Table of Contents

1. Introduction -- 2. Subjects and materials -- 3. Methods -- 4. Reults -- 5. Discussion -- Appendices -- Abbreviations -- References.


Bibliography: pages 121-134 Empirical thesis.

Awarding Institution

Macquarie University

Degree Type

Thesis MRes


MRes, Macquarie University, Faculty of Medicine and Health Sciences, Department of Biomedical Sciences

Department, Centre or School

Department of Biomedical Sciences

Year of Award


Principal Supervisor

Ian Blair

Additional Supervisor 1

Jennifer Ann Fifita

Additional Supervisor 2

Shu Yang


Copyright Sandrine Kim Kiow Chan Moi Fat 2019. Copyright disclaimer:




1 online resource (xviii, 134 pages) colour illustrations

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