title: Sequence-specific modification of a beta-thalassemia locus by small DNA fragments in human erythroid progenitor cells.
abstract: Gene therapy has been proposed as a definitive cure of beta-thalassemia. We applied a gene targeting approach, based on the introduction of small DNA fragments (SDF) into erythroid progenitor cells, to specifically modify the beta-globin gene sequence at codon 39. The strategy was first tested in normal individuals by delivering mutant SDF that were able to produce the beta-39 (C->T) mutation. Secondly, wild-type SDF were electroporated into target cells of beta-3i9/beta-39 b-thalassemic patients to correct the endogenous mutation. In both cases, gene modification was assayed by allele-specific polymerase chain reaction of DNA and mRNA, by restriction fragment length polymorphism analysis and by direct sequencing.
authors.forename: Alessia
authors.initials: A
authors.lastname: Colosimo
authors.forename: Valentina
authors.initials: V
authors.lastname: Guida
authors.forename: Ivana
authors.initials: I
authors.lastname: Antonucci
authors.forename: Tiziana
authors.initials: T
authors.lastname: Bonfini
authors.forename: Liborio
authors.initials: L
authors.lastname: Stuppia
authors.forename: Bruno
authors.initials: B
authors.lastname: Dallapiccola
mesh_terms: D000483:Alleles; D001483:Base Sequence; D003062:Codon; D053938:DNA Fragmentation; D018274:Electroporation; D015672:Erythroid Precursor Cells / Q000166:cytology*; D018390:Gene Targeting; D005914:Globins / Q000235:genetics; D006579:Heterozygote; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D017086:beta-Thalassemia / Q000235:genetics*
chemical_list: D003062:Codon; D012333:RNA, Messenger; D005914:Globins