Polyamides containing N-methylpyrrole and N-methylimidazole bind predetermined DNA sequences with affinity and specificity comparable to natural DNA binding proteins. They are cell permeable and able to inhibit gene transcription in vivo. Yet there is no report on how to use these polyamides to activate gene transcription. It is proposed here that FK506-linked polyamides are very likely to be able to target FKBP-VP16 transcription activation domain fusion protein (FKBPVP16) to DNA and activate gene transcription. Therefore, FK506-linked polyamide adapter molecules will be synthesized. Their ability to recruit FKBPVP16 to target DNA and transcription activity of the polyamide-FKBPVP16 complex will be tested in collaboration with Ron Prywes lab at Columbia University. Since dimerized transcription factors can bind longer DNA sequences with higher affinity than monomers, the difference between monomeric and dimeric artificial transcription factors in their transcriptional activity will be investigated. If this system works well, it will be the first example of artificial transcription factors that has the potential to activate the transcription of whatever gene, including the genes that play key roles in cancers. These artificial transcription factors will be powerful tools for biomedical research and have the potential to be used directly in gene therapy.
