Abstract
Mononuclear cells from peripheral blood of thalassemic patients were treated with morpholino oligonucleotides antisense to aberrant splice sites in mutant beta-globin precursor mRNAs (pre-mRNAs). The oligonucleotides restored correct splicing and translation of beta-globin mRNA, increasing the hemoglobin (Hb) A synthesis in erythroid cells from patients with IVS2-654/beta(E), IVS2-745/IVS2-745, and IVS2-745/IVS2-1 genotypes. The maximal Hb A level for repaired IVS2-745 mutation was approximately 30% of normal; Hb A was still detectable 9 days after a single treatment with oligonucleotide. Thus, expression of defective beta-globin genes was repaired and significant level of Hb A was restored in a cell population that would be targeted in clinical applications of this approach.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Cell Nucleus / genetics
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Cells, Cultured
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Erythrocytes / metabolism*
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Erythroid Precursor Cells / metabolism
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Fluorescent Antibody Technique
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Genetic Therapy*
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Globins / genetics
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Hemoglobin A / biosynthesis*
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Hemoglobin A / genetics*
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Humans
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Mutation / genetics
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Oligonucleotides, Antisense / genetics
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Oligonucleotides, Antisense / therapeutic use
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RNA Precursors / genetics
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RNA Precursors / metabolism
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RNA Splicing / genetics
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Spliceosomes / genetics
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Time Factors
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beta-Thalassemia / blood*
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beta-Thalassemia / genetics
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beta-Thalassemia / therapy*
Substances
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Oligonucleotides, Antisense
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RNA Precursors
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RNA, Messenger
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Globins
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Hemoglobin A