Haematologica (Jan 2007)

Sequence-specific modification of a β-thalassemia locus by small DNA fragments in human erythroid progenitor cells

  • Alessia Colosimo,
  • Valentina Guida,
  • Ivana Antonucci,
  • Tiziana Bonfini,
  • Liborio Stuppia,
  • Bruno Dallapiccola

DOI
https://doi.org/10.3324/haematol.10560
Journal volume & issue
Vol. 92, no. 1

Abstract

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Gene therapy has been proposed as a definitive cure for β-thalassemia. We applied a gene targeting approach, based on the introduction of small DNA fragments (SDF) into erythroid progenitor cells, to specifically modify the β-globin gene sequence at codon 39. The strategy was first tested in normal individuals by delivering mutant SDF that were able to produce the β39 (C→T) mutation. Secondly, wild-type SDF were electroporated into target cells of β39/β39. β-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.