PNA as a potential modulator of COL7A1 gene expression in dominant dystrophic epidermolysis bullosa: a physico-chemical study
PNA as a potential modulator of COL7A1 gene expression in dominant dystrophic epidermolysis bullosa: a physico-chemical study
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DOI：
10.1039/c3mb70283a
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发表时间：
2013-01-01
影响因子：
--
通讯作者：
Giancola, Concetta
中科院分区：
文献类型：
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作者：
Amato, Jussara;Stellato, Marco Ignazio;Giancola, Concetta
显性疾病是发生在杂合子状态的单基因疾病。突变的等位基因发挥显性作用，因为它产生一种异常多肽，干扰正常等位基因产物的功能。肽核酸(PNA)通过选择性地沉默携带显性突变的等位基因，为治疗显性疾病提供了一条潜在的途径。在这里，我们已经合成并研究了与C.5272-38T>；A序列变异完全互补的15聚体PNA的性质，该序列变异鉴定了导致显性营养不良性大疱性表皮松解症(DDEB)的一个重复突变的COL7A1等位基因，这是一种孟德尔疾病，以皮肤起泡为特征。PNA在C端与四个赖氨酸残基连接，在N端与一个荧光探针连接。理化结果证明，在体外可以形成稳定的、选择性的PNA/突变体-DNA异源双链。有趣的是，当将PNA导入正常的人成纤维细胞后，PNA正确地定位在细胞核中。我们的结果为DDEB患者打开了新的治疗可能性。
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.