title: PNA as a potential modulator of COL7A1 gene expression in dominant dystrophic epidermolysis bullosa: a physico-chemical study.
abstract: Dominant diseases are single gene disorders occurring in the heterozygous state. The mutated allele exerts a dominant effect because it produces an abnormal polypeptide that interferes with the function of the normal allele product. Peptide Nucleic Acids (PNAs) offer a route for a potential therapy for dominant diseases by selectively silencing the allele carrying the dominant mutation. Here, we have synthesized and studied the properties of a 15-mer PNA fully complementary to the site of the c.5272-38T>A sequence variation, which identifies a recurrent mutant COL7A1 allele causing dominant dystrophic epidermolysis bullosa (DDEB), a mendelian disease characterized by skin blistering. The PNA was conjugated with four lysine residues at the C-terminus and a fluorescent probe at the N-terminus. Physico-chemical results proved the formation of a stable, selective PNA/mutant-DNA heteroduplex in vitro. Intriguingly, when transfected into normal human fibroblasts, the PNA correctly localized in the cell nucleus. Our results open new therapeutic possibilities for patients with DDEB.
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", via D. Montesano 49, 80131 Naples, Italy. giancola@unina.it.
authors.forename: Jussara
authors.initials: J
authors.lastname: Amato
authors.forename: Marco Ignazio
authors.initials: MI
authors.lastname: Stellato
authors.forename: Elio
authors.initials: E
authors.lastname: Pizzo
authors.forename: Luigi
authors.initials: L
authors.lastname: Petraccone
authors.forename: Giorgia
authors.initials: G
authors.lastname: Oliviero
authors.forename: Nicola
authors.initials: N
authors.lastname: Borbone
authors.forename: Gennaro
authors.initials: G
authors.lastname: Piccialli
authors.forename: Angela
authors.initials: A
authors.lastname: Orecchia
authors.forename: Barbara
authors.initials: B
authors.lastname: Bellei
authors.forename: Daniele
authors.initials: D
authors.lastname: Castiglia
authors.forename: Concetta
authors.initials: C
authors.lastname: Giancola
mesh_terms: D002152:Calorimetry, Differential Scanning; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002942:Circular Dichroism; D024143:Collagen Type VII / Q000235:genetics* / Q000378:metabolism*; D016108:Epidermolysis Bullosa Dystrophica / Q000235:genetics*; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000378:metabolism*
chemical_list: C569053:COL7A1 protein, human; D024143:Collagen Type VII; D009692:Nucleic Acid Heteroduplexes; D020135:Peptide Nucleic Acids