Barre F X, Giovannangeli C, Hélène C, Harel-Bellan A
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.
Nucleic Acids Res. 1999 Feb 1;27(3):743-9. doi: 10.1093/nar/27.3.743.
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.
三链螺旋形成寡核苷酸（TFOs）是人工调节基因活性的潜在有力工具。特别是，它们可用于将损伤特异性引入选定的靶序列：通过与补骨脂素偶联的TFOs可引入链间交联和单加合物。这些策略的效率取决于细胞修复这些损伤的能力，而这一问题仍存在争议。在此，我们使用与补骨脂素偶联的TFOs并以酵母作为方便的细胞测试系统，表明链间交联的修复在数量上较差，从而导致靶基因活性的有效改变。此外，这些损伤在高比例的细胞中导致突变的引入。我们表明，这些突变是由易错修复途径单独或与核苷酸切除修复共同产生的。综上所述，这些结果表明，与补骨脂素偶联的TFOs可用于高效地使基因失活。
