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Modelling Inherited Prion Diseases In Mice

Owusu-Yaw, Bernie Simone; (2022) Modelling Inherited Prion Diseases In Mice. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

Prion diseases are fatal neurodegenerative diseases of mammals, caused by a misfolded infectious protein. There are 3 aetiological forms namely sporadic, acquired or inherited. The genetic component makes it possible for transgenic technology to be applied to the modelling of prion diseases. In 2013 Mead et al. reported a novel prion disease linked to a stop codon mutation at residue 163 (Y163X) in the human prion protein gene (PRNP). This mutation results in the production of a truncated prion protein (PrP) lacking both glycosylation sites and GPI anchor, and it is associated with an atypical prion disease phenotype including diarrhoea, autonomic neuropathy and the accumulation of PrP amyloid in peripheral organs and blood vessels. Typically, prion diseases predominantly affect the nervous system so the uniqueness of the Y163X disease phenotype makes it a good candidate for the study of prion therapeutics. The aim of this project was therefore to develop mouse models expressing human PrP Y163X that recapitulate human disease and then to study ageing cohorts of homozygous and heterozygous mice for spontaneous amyloid deposition. Two different approaches were pursued as follows: (1) Conventional transgenic approach (2) Gene targeted approach (CRISPR- Cas9 and knock-in technology). Six transgenic lines expressing HuPrP 163X under the control of the ubiquitous CAG promoter were generated alongside transgenic wild-type controls expressing wild-type human PrP. Y163X knock-in mice and control knock-in mice expressing wild-type human PrP generated from blastocyst injection of embryonic stem cells were made commercially. These lines were characterised for expression of the truncated 163X human prion protein using techniques such as western blotting, ELISA, fluorescent immunohistochemistry and RT-PCR. Aging cohorts from homozygous lines were set up in long term observation experiments. A panel of CNS and peripheral tissues has been isolated at defined time points for immunohistochemical analysis to determine if the expression of HuPrP Y163X in transgenic and knock-in mice is sufficient to cause spontaneous PrP amyloid deposition.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: Modelling Inherited Prion Diseases In Mice
Language: English
Additional information: Copyright © The Author 2021. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request.
UCL classification: UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL EGA Institute for Womens Health
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Institute of Prion Diseases > MRC Prion Unit at UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL EGA Institute for Womens Health > Maternal and Fetal Medicine
UCL
URI: https://discovery.ucl.ac.uk/id/eprint/10147141
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