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• W2005735769 abstract "A single nucleotide polymorphism (SNP) in the sickle β-globin gene (β S ) leads to sickle cell anemia. Sickling increases sharply with deoxy sickle Hb concentration and decreases with increasing fetal γ-globin concentration. Measures that decrease sickle Hb concentration should have an antisickling effect. RNA interference (RNAi) uses small interfering (si)RNAs for sequence-specific gene silencing. A β S siRNA with position 10 of the guide strand designed to align with the targeted β S SNP specifically silences β S gene expression without affecting the expression of the γ-globin or normal β-globin (β A ) genes. Silencing is increased by altering the 5′ end of the siRNA antisense (guide) strand to enhance its binding to the RNA-induced silencing complex (RISC). Specific β S silencing was demonstrated by using a luciferase reporter and full-length β S cDNA transfected into HeLa cells and mouse erythroleukemia cells, where it was expressed in the context of the endogenous β-globin gene promoter and the locus control region enhancers. When this strategy was used to target β E , silencing was not limited to the mutant gene but also targeted the normal β A gene. siRNAs, mismatched with their target at position 10, guided mRNA cleavage in all cases except when two bulky purines were aligned. The specific silencing of the β S -globin gene, as compared with β E , as well as studies of silencing SNP mutants in other diseases, indicates that siRNAs developed to target a disease-causing SNP will be specific if the mutant residue is a pyrimidine and the normal residue is a purine." @default.
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• W2005735769 date "2006-04-11" @default.
• W2005735769 modified "2023-10-15" @default.
• W2005735769 title "Determinants of specific RNA interference-mediated silencing of human β-globin alleles differing by a single nucleotide polymorphism" @default.
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• W2005735769 doi "https://doi.org/10.1073/pnas.0601309103" @default.
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