TY - JOUR
T1 - PNA as a potential modulator of COL7A1 gene expression in dominant dystrophic epidermolysis bullosa
T2 - A physico-chemical study
AU - Amato, Jussara
AU - Stellato, Marco Ignazio
AU - Pizzo, Elio
AU - Petraccone, Luigi
AU - Oliviero, Giorgia
AU - Borbone, Nicola
AU - Piccialli, Gennaro
AU - Orecchia, Angela
AU - Bellei, Barbara
AU - Castiglia, Daniele
AU - Giancola, Concetta
PY - 2013/12
Y1 - 2013/12
N2 - 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.
AB - 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.
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U2 - 10.1039/c3mb70283a
DO - 10.1039/c3mb70283a
M3 - Article
C2 - 24121392
AN - SCOPUS:84887030996
VL - 9
SP - 3166
EP - 3174
JO - Molecular BioSystems
JF - Molecular BioSystems
SN - 1742-206X
IS - 12
ER -