Tyrosinase gene correction using fluorescent oligonucleotides

Pigment Cell Res. 2003 Apr;16(2):133-8. doi: 10.1034/j.1600-0749.2003.00021.x.

Abstract

Gene therapy and production of mutated cell lines or animal models should be improved significantly once efficient controlled gene targeting strategies are developed. We used short single-stranded oligodeoxynucleotides (ODN), in some cases coupled to the fluorescent dye fluorescein isothiocyanate (FITC), to correct an endogenic natural point mutation in melanocytes in culture. The addition of the FITC molecule to the 5' extremity of the ODN did not interfere with the efficiency of the reversion of the mutation and did not have any deleterious side-effects. The use of fluorescent ODN could lead to great improvement in the technique. In particular, it may facilitate sorting of the transfected cells in the treated population, and thereby significantly increase the percentage of corrected cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / chemistry
  • Cell Line
  • Cell Separation / methods
  • Fluorescein-5-isothiocyanate / pharmacology*
  • Fluorescein-5-isothiocyanate / therapeutic use
  • Gene Targeting / methods*
  • Genetic Therapy / methods*
  • Humans
  • Monophenol Monooxygenase / genetics*
  • Oligodeoxyribonucleotides / pharmacology*
  • Oligodeoxyribonucleotides / therapeutic use
  • Point Mutation / drug effects*
  • Point Mutation / genetics
  • Transfection / methods

Substances

  • Biomarkers
  • Oligodeoxyribonucleotides
  • Monophenol Monooxygenase
  • Fluorescein-5-isothiocyanate