Tau aggregation and propagation are key features of Alzheimer’s disease and other tauopathies. In this project, we will further characterize tau-specific antibodies reported to recognize low-n tau oligomers, with the aim of mapping their epitopes within the tau repeat domain (RD).
The student (f/m/x) will express different tau RD deletion constructs, such as variants lacking individual repeat regions, in HEK cells and assess antibody binding by Western blotting and immunofluorescence. If time permits, selected regions will be further analyzed by mutagenesis for finer epitope mapping.
In parallel, the same tau RD constructs will be tested for their ability to undergo seed-induced aggregation following exposure to exogenous tau seeds. This will initially be assessed using sedimentation assays. If time permits, stable cell lines expressing selected constructs may be generated by lentiviral transduction for image-based tau seeding assays.
Methods and techniques the student will learn:
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Mammalian cell culture of HEK cells
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Transient transfection of tau RD constructs
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Optional: lentiviral transduction and generation of stable cell populations
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Western blotting for tau construct expression and antibody binding
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Immunofluorescence staining and confocal microscopy
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Epitope mapping using deletion constructs
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Optional: site-directed mutagenesis for finer epitope mapping
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Biochemical sedimentation assays to separate soluble and insoluble tau fractions
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Image-based aggregation assays and quantification, if stable cell lines are generated
