Designing a high-throughput screen for small molecule modulators of the molecular motor KIF1A

Author: 
Date created: 
2020-12-07
Identifier: 
etd21161
Keywords: 
KIF1A
Neuronal Transport
Neurodegenerative Disorders
Assay Development
High-Throughput Screen
Drug Discovery
Abstract: 

The neuron-specific kinesin, KIF1A, is involved in the transport of cargos critical for neuronal function such as synaptic vesicle precursors and dense-core vesicles. Notably, mutations in KIF1A are implicated in the pathogenesis of neurological disorders such as hereditary spastic paraplegia. Therapies that specifically target KIF1A do not currently exist. I developed a high-throughput, phenotypic screen for small molecule modulators of KIF1A. I used two-color fluorescent imaging to identify a cell-based, phenotypic assay that allows for characterization of the inactive, autoinhibited form of KIF1A and the active form of KIF1A. When expressed in COS-7 and SK-N-SH cells, wild-type KIF1A-GFP is diffusely distributed and does not co-localize with the microtubule marker EB3-mKate2. The hyperactive mutant V8M KIF1A-GFP does co-localize with microtubules and distributes peripherally. These findings support the feasibility of a functional assay to distinguish between the active and inactive forms of KIF1A in a high-throughput screening format.

Document type: 
Thesis
Rights: 
This thesis may be printed or downloaded for non-commercial research and scholarly purposes. Copyright remains with the author.
File(s): 
Supervisor(s): 
Michael Silverman
Department: 
Science: Department of Molecular Biology and Biochemistry
Thesis type: 
(Thesis) M.Sc.
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