title: Inhibiting transcription of chromosomal DNA with antigene peptide nucleic acids.
abstract: Synthetic molecules that recognize specific sequences within cellular DNA are potentially powerful tools for investigating chromosome structure and function. Here, we designed antigene peptide nucleic acids (agPNAs) to target the transcriptional start sites for the human progesterone receptor B (hPR-B) and A (hPR-A) isoforms at sequences predicted to be single-stranded within the open complex of chromosomal DNA. We found that the agPNAs were potent inhibitors of transcription, showing for the first time that synthetic molecules can recognize transcription start sites inside cells. Breast cancer cells treated with agPNAs showed marked changes in morphology and an unexpected relationship between the strictly regulated levels of hPR-B and hPR-A. We confirmed these phenotypes using siRNAs and antisense PNAs, demonstrating the power of combining antigene and antisense strategies for gene silencing. agPNAs provide a general approach for controlling transcription initiation and a distinct option for target validation and therapeutic development.
authors.affiliation: Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390, USA.
authors.forename: Bethany A
authors.initials: BA
authors.lastname: Janowski
authors.forename: Kunihiro
authors.initials: K
authors.lastname: Kaihatsu
authors.forename: Kenneth E
authors.initials: KE
authors.lastname: Huffman
authors.forename: Jacob C
authors.initials: JC
authors.lastname: Schwartz
authors.forename: Rosalyn
authors.initials: R
authors.lastname: Ram
authors.forename: Daniel
authors.initials: D
authors.lastname: Hardy
authors.forename: Carole R
authors.initials: CR
authors.lastname: Mendelson
authors.forename: David R
authors.initials: DR
authors.lastname: Corey
mesh_terms: D002877:Chromosomes, Human; D003598:Cytoskeletal Proteins; D004247:DNA / Q000378:metabolism*; D020868:Gene Silencing; D006801:Humans; D008958:Models, Molecular; D020135:Peptide Nucleic Acids / Q000378:metabolism / Q000494:pharmacology*; D010750:Phosphoproteins / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism / Q000494:pharmacology; D011980:Receptors, Progesterone / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D024363:Transcription Initiation Site / Q000502:physiology*; D014158:Transcription, Genetic / Q000187:drug effects*; D000099149:Ezrin
chemical_list: D003598:Cytoskeletal Proteins; D004247:DNA; D020135:Peptide Nucleic Acids; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D011980:Receptors, Progesterone; D000099149:Ezrin