Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.774288
Title: Investigating the molecular causes of inherited thrombocytopenia using CRISPR-Cas9 gene editing and super-resolution microscopy
Author: Khan, Abdullah O.
ISNI:       0000 0004 7961 4944
Awarding Body: University of Birmingham
Current Institution: University of Birmingham
Date of Award: 2019
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Abstract:
Inherited thrombocytopenias (ITs) are a heterogenous group of disorders characterised by an abnormal platelet count (< 150 x 109/L). Functional investigations into the mechanistic causes of ITs often suffer from numerous technical limitations, including a lack of patient material and the absence of platelet producing cell lines. In this thesis, protocols to address these issues through the application of CRISPR (Clustered Regularly Interspaced Palindromic Repeats) genome editing and stem cell differentiation are developed and applied to the study of the platelet and megakaryocyte specific β-tubulin isoform encoded by the TubB1 gene. First, a novel method of single molecule super-resolution microscopy through the CRISPR mediated editing of cell lines is devised, and attempts to transfer this approach to stem cells are currently underway. Secondly, existing methods for the differentiation of induced pluripotent stem cells into megakaryocytes are adapted to generate mature, proplatelet forming cells for in vitro phenotypic screening. Finally, two families from the UK GAPP (Genotyping and Phenotyping of Platelets) study with TubB1 mutations affecting C-terminal function are investigated to determine the role of C-terminal post-translational modifications (PTMs) on ITs. Two novel PTMs (polyglutamylation and polyglycylation) which regulate motor protein processivity in both platelet production and function are reported.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.774288  DOI: Not available
Keywords: QH426 Genetics
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