Abstract
[Truncated] The alpha (α) thalassemias are a group of disorders occurring as a result of decreased synthesis of α-globin chains due to deletions of α-globin genes or, less commonly, point mutations. The main aim of this research was to elucidate molecular mechanisms and events regulating human α-globin production in an in vitro system and perform comparative analysis between normal and α-thalassemia subtypes. Research was undertaken to investigate the impact of non-deletional α-thalassemia, caused mainly by point mutations, on various steps involved in α-globin expression with focus on pretranscriptional
and transcriptional processes.
The need for an in-vitro system to analyse the pathological effects of various mutations wasidentified during the course of routine diagnostic work for the investigation of patients withpotential α-thalassemia. In cases where the common alpha globin gene deletions have been excluded, sequencing of the alpha1 (HBA1) and alpha2 (HBA2) genes is undertaken and this has yielded novel base substitutions in some cases. In order to provide information to the requesting clinician regarding whether the base substitution is pathogenic or polymorphic it was necessary to develop a system to investigate the impact of the mutation
on α-globin expression. Therefore, this study resulted in developing an in vitro system with expression vectors carrying the wild type α2- and α1-globin genes as well as various novel mutations. The generated system is driven by the α-globin gene’s own regulatory elements present at the 5` and 3` UTRs rather than recombinant elements such as viral promoter and polyadenylation signals. This gave us an advantage of studying mutations located at these regions as well as exonic and intronic mutations.
and transcriptional processes.
The need for an in-vitro system to analyse the pathological effects of various mutations wasidentified during the course of routine diagnostic work for the investigation of patients withpotential α-thalassemia. In cases where the common alpha globin gene deletions have been excluded, sequencing of the alpha1 (HBA1) and alpha2 (HBA2) genes is undertaken and this has yielded novel base substitutions in some cases. In order to provide information to the requesting clinician regarding whether the base substitution is pathogenic or polymorphic it was necessary to develop a system to investigate the impact of the mutation
on α-globin expression. Therefore, this study resulted in developing an in vitro system with expression vectors carrying the wild type α2- and α1-globin genes as well as various novel mutations. The generated system is driven by the α-globin gene’s own regulatory elements present at the 5` and 3` UTRs rather than recombinant elements such as viral promoter and polyadenylation signals. This gave us an advantage of studying mutations located at these regions as well as exonic and intronic mutations.
| Original language | English |
|---|---|
| Qualification | Doctor of Philosophy |
| Publication status | Unpublished - 2014 |