Genome editing in K562 cells suggests a functional role for the XmnI Gg polymorphism: a widely used genetic marker in ß-thalassemia and sickle cell disease patients.
Cell Mol Biol (Noisy-le-grand)
; 70(7): 230-236, 2024 Jul 28.
Article
em En
| MEDLINE
| ID: mdl-39097869
ABSTRACT
The XmnI Gg -158 C/T polymorphism has been widely associated with fetal hemoglobin (HbF) levels, the severity of disease, and the response to the drug hydroxyurea (HU) in both ß-thalassemia (ß-thal) and sickle cell disease (SCD) patients. However, the functional significance of this single nucleotide polymorphism (SNP) remains unclear. To gain insight, green fluorescence protein (GFP) cassettes harboring the XmnI C or T alleles in their left homology arms (i.e. Gg promoters) were knocked into the Gg gene(s) of K562 cells via CRISPR/Cas9. Subsequently, the GFP fluorescence levels were compared in the ensuing cell populations and isolated clones. In both instances, median fluorescence intensities (MFI) of the knockin cells having the inserted XmnI T allele were higher than those having the XmnI C allele. Our results suggest that the XmnI T allele can increase Gg expression in K562 cells. The possible functional significance of the XmnI Gg -158 C/T polymorphism provides a rationale for the aforementioned associations. Furthermore, the XmnI polymorphism as a functional SNP substantiates its importance as a prognostic marker.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Talassemia beta
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Polimorfismo de Nucleotídeo Único
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Alelos
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Sistemas CRISPR-Cas
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Edição de Genes
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Anemia Falciforme
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article