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Nucleic Acids Research, Vol 27, Issue 3 743-749, Copyright © 1999 by Oxford University Press

ARTICLES

Covalent crosslinks introduced via a triple helix-forming oligonucleotide coupled to psoralen are inefficiently repaired

FX Barre, C Giovannangeli, C Helene and A Harel-Bellan
CNRS UPR 9079, Institut de Recherche sur le Cancer, 7 rue Guy Moquet, 94801 Villejuif, France andLaboratoire de Biophysique, INSERM U 201, CNRS URA481, MHN, Paris, France.

Triple helix-forming oligonucleotides (TFOs) represent potentially powerful tools to artificially modulate gene activity. In particular, they can be used to specifically introduce a lesion into a selected target sequence: interstrand crosslinks and monoadducts can be introduced via TFOs coupled to psoralen. The efficiency of these strategies depends on the cell ability to repair these lesions, an issue which is still controversial. Here we show, using psoralen- coupled TFOs and the yeast as a convenient cellular test system, that interstrand crosslinks are quantitatively poorly repaired, resulting in an efficient modification of target gene activity. In addition, these lesions result in the introduction of mutations in a high proportion of cells. We show that these mutations are generated by the Error-Prone Repair pathway, alone or in combination with Nucleotide Excision Repair. Taken together, these results suggest that TFOs coupled to psoralen could be used to inactivate a gene with significant efficiency.
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