Tyrosinase gene correction using fluorescent oligonucleotides.
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.
Biet
Elodie
E
Recombination and Genomic Instability, UMR 2027 CNRS-Institut Curie, Orsay, France.
Alberti
Christophe
C
Faccella
Paolo
P
Sun
Jian Sheng
JS
Dutreix
Marie
M
Larue
Lionel
L
D000818:Animals; D015415:Biomarkers / Q000737:chemistry; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D016650:Fluorescein-5-isothiocyanate / Q000494:pharmacology* / Q000627:therapeutic use; D018390:Gene Targeting / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D014442:Monophenol Monooxygenase / Q000235:genetics*; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology* / Q000627:therapeutic use; D017354:Point Mutation / Q000187:drug effects* / Q000235:genetics; D014162:Transfection / Q000379:methods
D015415:Biomarkers; D009838:Oligodeoxyribonucleotides; D014442:Monophenol Monooxygenase; D016650:Fluorescein-5-isothiocyanate