Zinc-finger nuclease-mediated correction of α-thalassemia in iPS cells

Blood. 2012 Nov 8;120(19):3906-14. doi: 10.1182/blood-2012-03-420703. Epub 2012 Sep 21.

Abstract

Induced pluripotent stem (iPS) cell technology holds vast promises for a cure to the hemoglobinopathies. Constructs and methods to safely insert therapeutic genes to correct the genetic defect need to be developed. Site-specific insertion is a very attractive method for gene therapy because the risks of insertional mutagenesis are eliminated provided that a "safe harbor" is identified, and because a single set of validated constructs can be used to correct a large variety of mutations simplifying eventual clinical use. We report here the correction of α-thalassemia major hydrops fetalis in transgene-free iPS cells using zinc finger-mediated insertion of a globin transgene in the AAVS1 site on human chromosome 19. Homozygous insertion of the best of the 4 constructs tested led to complete correction of globin chain imbalance in erythroid cells differentiated from the corrected iPS cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation
  • Cell Line
  • Endonucleases / metabolism*
  • Erythroblasts / cytology
  • Erythroblasts / metabolism
  • Erythrocyte Indices
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Gene Expression Regulation
  • Gene Order
  • Homozygote
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism*
  • Recombination, Genetic
  • Transgenes
  • Zinc Fingers*
  • alpha-Globins / genetics
  • alpha-Globins / metabolism
  • alpha-Thalassemia / genetics
  • alpha-Thalassemia / metabolism*
  • alpha-Thalassemia / therapy

Substances

  • alpha-Globins
  • Endonucleases