[HTML][HTML] Repair of a splicing defect in erythroid cells from patients with β-thalassemia/HbE disorder

T Suwanmanee, H Sierakowska, S Fucharoen, R Kole - Molecular Therapy, 2002 - cell.com
T Suwanmanee, H Sierakowska, S Fucharoen, R Kole
Molecular Therapy, 2002cell.com
A HeLa cell line stably expressing the human β-globin gene carrying thalassemic mutations
β E/IVS1-6 served as a thalassemia model for repair of aberrant splicing of β E-globin pre-
mRNA with antisense oligonucleotides. Treatment of β E/IVS1-6 HeLa cells with a
morpholino oligonucleotide targeted immediately upstream of the aberrant 5′ splice site
activated by the mutations resulted in an increase in the amount of correctly spliced β E-
globin mRNA in a dose-dependent and sequence-specific fashion. The repaired β E-globin …
Abstract
A HeLa cell line stably expressing the human β-globin gene carrying thalassemic mutations βE/IVS1-6 served as a thalassemia model for repair of aberrant splicing of βE-globin pre-mRNA with antisense oligonucleotides. Treatment of βE/IVS1-6 HeLa cells with a morpholino oligonucleotide targeted immediately upstream of the aberrant 5′ splice site activated by the mutations resulted in an increase in the amount of correctly spliced βE-globin mRNA in a dose-dependent and sequence-specific fashion. The repaired βE-globin mRNA was stable and could be translated into full-length βE-globin polypeptide. Application of the same oligonucleotide to erythroid progenitor cells from two β-thalassemia/HbE patients resulted in an approximately 70% increase in correct βE-globin mRNA and 36% increase in hemoglobin E. The erythroid progenitor cells represent the actual targets for the clinical application of antisense repair of defective pre-mRNAs.
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