Direct correction of haemoglobin E β-thalassaemia using base editors
Haemoglobin E (HbE) β-thalassaemia causes approximately 50% of all severe thalassaemia worldwide; equating to around 30,000 births per year. HbE β-thalassaemia is due to a point mutation in codon 26 of the human HBB gene on one allele (GAG; glutamatic acid → AAG; lysine, E26K), and any mutation caus...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Springer Nature
2023
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| _version_ | 1797110326882205696 |
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| author | Badat, M Ejaz, A Hua, P Rice, S Zhang, W Hentges, LD Fisher, CA Denny, N Schwessinger, R Yasara, N Roy, NBA Issa, F Roy, A Telfer, P Hughes, J Mettananda, S Higgs, DR Davies, JOJ |
| author_facet | Badat, M Ejaz, A Hua, P Rice, S Zhang, W Hentges, LD Fisher, CA Denny, N Schwessinger, R Yasara, N Roy, NBA Issa, F Roy, A Telfer, P Hughes, J Mettananda, S Higgs, DR Davies, JOJ |
| author_sort | Badat, M |
| collection | OXFORD |
| description | Haemoglobin E (HbE) β-thalassaemia causes approximately 50% of all severe thalassaemia worldwide; equating to around 30,000 births per year. HbE β-thalassaemia is due to a point mutation in codon 26 of the human HBB gene on one allele (GAG; glutamatic acid → AAG; lysine, E26K), and any mutation causing severe β-thalassaemia on the other. When inherited together in compound heterozygosity these mutations can cause a severe thalassaemic phenotype. However, if only one allele is mutated individuals are carriers for the respective mutation and have an asymptomatic phenotype (β-thalassaemia trait). Here we describe a base editing strategy which corrects the HbE mutation either to wildtype (WT) or a normal variant haemoglobin (E26G) known as Hb Aubenas and thereby recreates the asymptomatic trait phenotype. We have achieved editing efficiencies in excess of 90% in primary human CD34 + cells. We demonstrate editing of long-term repopulating haematopoietic stem cells (LT-HSCs) using serial xenotransplantation in NSG mice. We have profiled the off-target effects using a combination of circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-seq) and deep targeted capture and have developed machine-learning based methods to predict functional effects of candidate off-target mutations. |
| first_indexed | 2024-03-07T07:53:25Z |
| format | Journal article |
| id | oxford-uuid:b9f92570-f238-4634-881c-8ac14f957fba |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:53:25Z |
| publishDate | 2023 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | oxford-uuid:b9f92570-f238-4634-881c-8ac14f957fba2023-08-02T16:25:34ZDirect correction of haemoglobin E β-thalassaemia using base editorsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:b9f92570-f238-4634-881c-8ac14f957fbaEnglishSymplectic ElementsSpringer Nature2023Badat, MEjaz, AHua, PRice, SZhang, WHentges, LDFisher, CADenny, NSchwessinger, RYasara, NRoy, NBAIssa, FRoy, ATelfer, PHughes, JMettananda, SHiggs, DRDavies, JOJHaemoglobin E (HbE) β-thalassaemia causes approximately 50% of all severe thalassaemia worldwide; equating to around 30,000 births per year. HbE β-thalassaemia is due to a point mutation in codon 26 of the human HBB gene on one allele (GAG; glutamatic acid → AAG; lysine, E26K), and any mutation causing severe β-thalassaemia on the other. When inherited together in compound heterozygosity these mutations can cause a severe thalassaemic phenotype. However, if only one allele is mutated individuals are carriers for the respective mutation and have an asymptomatic phenotype (β-thalassaemia trait). Here we describe a base editing strategy which corrects the HbE mutation either to wildtype (WT) or a normal variant haemoglobin (E26G) known as Hb Aubenas and thereby recreates the asymptomatic trait phenotype. We have achieved editing efficiencies in excess of 90% in primary human CD34 + cells. We demonstrate editing of long-term repopulating haematopoietic stem cells (LT-HSCs) using serial xenotransplantation in NSG mice. We have profiled the off-target effects using a combination of circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-seq) and deep targeted capture and have developed machine-learning based methods to predict functional effects of candidate off-target mutations. |
| spellingShingle | Badat, M Ejaz, A Hua, P Rice, S Zhang, W Hentges, LD Fisher, CA Denny, N Schwessinger, R Yasara, N Roy, NBA Issa, F Roy, A Telfer, P Hughes, J Mettananda, S Higgs, DR Davies, JOJ Direct correction of haemoglobin E β-thalassaemia using base editors |
| title | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_full | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_fullStr | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_full_unstemmed | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_short | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_sort | direct correction of haemoglobin e β thalassaemia using base editors |
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