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

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)129-130
Number of pages2
JournalHaematologica
Volume92
Issue number1
DOIs
Publication statusPublished - Jan 2007

Fingerprint

Erythroid Precursor Cells
Thalassemia
DNA
Mutation
Globins
Gene Targeting
Codon
Restriction Fragment Length Polymorphisms
Genetic Therapy
Genes
Alleles
Polymerase Chain Reaction
Messenger RNA

Keywords

  • β-thalassemia
  • Gene targeting
  • Gene therapy
  • HBB gene

ASJC Scopus subject areas

  • Hematology

Cite this

Sequence-specific modification of a β-thalassemia locus by small DNA fragments in human erythroid progenitor cells. / Colosimo, Alessia; Guida, Valentina; Antonucci, Ivana; Bonfini, Tiziana; Stuppia, Liborio; Dallapiccola, Bruno.

In: Haematologica, Vol. 92, No. 1, 01.2007, p. 129-130.

Research output: Contribution to journalArticle

Colosimo, Alessia ; Guida, Valentina ; Antonucci, Ivana ; Bonfini, Tiziana ; Stuppia, Liborio ; Dallapiccola, Bruno. / Sequence-specific modification of a β-thalassemia locus by small DNA fragments in human erythroid progenitor cells. In: Haematologica. 2007 ; Vol. 92, No. 1. pp. 129-130.
@article{dcd7ecece1364d1a921e8959e172c092,
title = "Sequence-specific modification of a β-thalassemia locus by small DNA fragments in human erythroid progenitor cells",
abstract = "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.",
keywords = "β-thalassemia, Gene targeting, Gene therapy, HBB gene",
author = "Alessia Colosimo and Valentina Guida and Ivana Antonucci and Tiziana Bonfini and Liborio Stuppia and Bruno Dallapiccola",
year = "2007",
month = "1",
doi = "10.3324/haematol.10560",
language = "English",
volume = "92",
pages = "129--130",
journal = "Haematologica",
issn = "0390-6078",
publisher = "NLM (Medline)",
number = "1",

}

TY - JOUR

T1 - Sequence-specific modification of a β-thalassemia locus by small DNA fragments in human erythroid progenitor cells

AU - Colosimo, Alessia

AU - Guida, Valentina

AU - Antonucci, Ivana

AU - Bonfini, Tiziana

AU - Stuppia, Liborio

AU - Dallapiccola, Bruno

PY - 2007/1

Y1 - 2007/1

N2 - 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.

AB - 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.

KW - β-thalassemia

KW - Gene targeting

KW - Gene therapy

KW - HBB gene

UR - http://www.scopus.com/inward/record.url?scp=33846923834&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846923834&partnerID=8YFLogxK

U2 - 10.3324/haematol.10560

DO - 10.3324/haematol.10560

M3 - Article

C2 - 17229648

AN - SCOPUS:33846923834

VL - 92

SP - 129

EP - 130

JO - Haematologica

JF - Haematologica

SN - 0390-6078

IS - 1

ER -