Gene therapy has been proposed as a definitive cure for ββ-thalassemia. We applied a gene targeting approach, based on the introduction of small DNA fragments (SDF) into erythroid progenitor cells, to specifically modify the ββ-globin gene sequence at codon 39. The strategy was first tested in normal individuals by delivering mutant SDF that were able to produce the ββ39 (C→→T) mutation. Secondly, wild-type SDF were electroporated into target cells of ββ39/ββ39. ββ-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.
Sequence-specific modification of a beta-thalassemia locus by small DNA fragments in human erythroid progenitor cells.
COLOSIMO, Alessia;ANTONUCCI, IVANA;BONFINI, Tiziana Donatella;STUPPIA, Liborio;
2007-01-01
Abstract
Gene therapy has been proposed as a definitive cure for ββ-thalassemia. We applied a gene targeting approach, based on the introduction of small DNA fragments (SDF) into erythroid progenitor cells, to specifically modify the ββ-globin gene sequence at codon 39. The strategy was first tested in normal individuals by delivering mutant SDF that were able to produce the ββ39 (C→→T) mutation. Secondly, wild-type SDF were electroporated into target cells of ββ39/ββ39. ββ-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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
