Recruitment of transcription factors to the target site by triplex-forming oligonucleotides.
Triplex-forming oligonucleotides (TFOs) are generally designed to inhibit transcription or DNA replication but can be used for more diverse purposes. Here we have designed a hairpin-TFO able to recruit transcription factors to a target DNA. The designed oligonucleotide contains a triplex-forming sequence, linked through a nucleotide loop to a double-stranded hairpin including the SRE enhancer of the c-fos gene promoter. We show here that this oligonucleotide can specifically recognise its DNA target at physiological salt and pH conditions. The stability of the triplex formed under these conditions is very high: >90% of the triplex remains intact after 24 h of incubation. Bound to the double-stranded target DNA, the oligonucleotide retains its ability to interact specifically with transcription factors, recruiting them to the proximity of the target DNA. Our results suggest that this type of oligonucleotide may prove useful in the design of new tools for artificial modulation of gene expression.
Svinarchuk
F
F
Laboratoire de Biochimie-Enzymologie, CNRS URA 147, Institut Gustave Roussy, 39 rue Camille Desmoulins, 94805 Villejuif Cedex, France. fedorsvi@igr.fr
Nagibneva
I
I
Cherny
D
D
Ait-Si-Ali
S
S
Pritchard
L L
LL
Robin
P
P
Malvy
C
C
Harel-Bellan
A
A
Chern
D
D
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism; D016762:Genes, fos; D015498:HIV-2 / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012191:Retroviridae Proteins / Q000235:genetics; D015302:Simian Immunodeficiency Virus / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism*; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics
D004279:DNA, Viral; D009841:Oligonucleotides; D012191:Retroviridae Proteins; D014157:Transcription Factors; C074286:VPX protein, Human immunodeficiency virus 2; C513059:VPX protein, Simian immunodeficiency virus; D054334:Viral Regulatory and Accessory Proteins; C051802:triplex DNA; D004247:DNA