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Title: | Differential expression of KLF1 in family studies and their role in globin gene switching |
Other Titles: | 10th Malta Medical School Conference : conference abstract book |
Authors: | Grech, Laura Cutajar, Jeremy Caruana, Mary Rose Scerri, Christian A. Galdies, Ruth Formosa, Robert Borg, Joseph J. Felice, Alex E. |
Keywords: | DNA-binding protein interactions Hemoglobin DNA -- Structure Proteins -- Structure Phenotype Erythrocytes |
Issue Date: | 2018 |
Publisher: | University of Malta. Medical School |
Citation: | Grech, L., Cutajar, J., Caruana, M.R., Scerri, C., Galdies, R. Formosa, R…. Felice, A. (2018). Differential expression of KLF1 in family studies and their role in globin gene switching. In P. Schembri-Wismayer, R. Galea, , C. Scerri, R. Muscat & A. Fenech (Eds.), 10th Malta Medical School Conference : conference abstract book (pp. 80). |
Abstract: | Introduction: Kruppel like factor 1 (KLF1) also known as the ‘Master regulator of erythropoiesis’ is located on the short arm of chromosome KLF1 activates a diverse set of genes that have an important role in the regulation of key pathways such as erythropoiesis, cell membrane and cytoskeleton. To date over 65 molecular variants have been recorded. Their haematological phenotypes range from the clinically unremarkable In(Lu) type of Lu(a-b-) blood group, variability in the HbA2 levels, congenital dyserythropoietic anaemia (CDA) and in most extreme cases hydrops foetalis secondary to profound anaemia. In 2010 sequencing revealed a nonsense mutation in KLF1 in a large Maltese family with hereditary persistence of foetal haemoglobin (HPFH). The p.K288X mutation was found to ablate the DNA binding domain of the key erythroid transcription factor. Methods: We explored the occurrence of additional KLF1 mutations with genotype – phenotype associations among a large number of cases, all from Malta with a borderline HbA2 but without beta globin gene mutations and other beta thalassaemia heterozygotes from the Malta Biobank and the Thalassaemia Clinic. Sequencing of the KLF1 gene was carried out. To study the promoter mutations in the KLF1 gene Dual- Luciferase reporter assays were performed on HEK293T cells and K562 cells transfected with the pGL4.10 vector containing either the wildtype promoter or the mutant promoter. This was followed by Electromobility Shift assays. Results: Four-hundred and twenty six subjects were collected. After sequencing of the KLF1 gene we identified 5 other families with the p.K288X mutation together with other nucleotide variants, six of which were in the KLF1 promoter. In both cell lines dual-luciferase assays showed a statistically significant difference between cells transfected with the wildtype KLF1 promoter and cells transfected with 7 different KLF1 promoter mutations. Electrophoretic Mobility Shift Assays on nuclear extracts further show DNA:Protein binding evidence and are currently being investigated Conclusion: This data further highlights the importance of KLF1 function in globin gene control and demonstrate the importance of KLF1 sequencing in patients with haematological features resembling beta-thalassaemia Disclosures: Dean’s Fund |
URI: | https://www.um.edu.mt/library/oar/handle/123456789/99416 |
ISSN: | 18133339 |
Appears in Collections: | Scholarly Works - FacHScABS |
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Differential_expression_of_KLF1_in_family_studies_and_their_role_in_globin_gene_switching.pdf | 120.09 kB | Adobe PDF | View/Open |
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