| Autor: |
Dal Mas, Andrea, Fortugno, Paola, Donadon, Irving, Levati, Lauretta, Castiglia, Daniele, Pagani, Franco |
| Zdroj: |
Human Mutation; May2015, Vol. 36 Issue 5, p504-512, 9p |
| Abstrakt: |
ABSTRACT The c.891 C> T synonymous transition in SPINK5 induces exon 11 ( E11) skipping and causes Netherton syndrome ( NS). Using a specific RNA-protein interaction assay followed by mass spectrometry analysis along with silencing and overexpression of splicing factors, we showed that this mutation affects an exonic bifunctional splicing regulatory element composed by two partially overlapping silencer and enhancer sequences, recognized by hn RNPA1 and Tra2β splicing factors, respectively. The C-to- T substitution concomitantly increases hn RNPA1 and weakens Tra2β-binding sites, leading to pathological E11 skipping. In hybrid minigenes, exon-specific U1 small nuclear RNAs ( Ex Spe U1s) that target by complementarity intronic sequences downstream of the donor splice site rescued the E11 skipping defect caused by the c.891 C> T mutation. Ex Spe U1 lentiviral-mediated transduction of primary NS keratinocytes from a patient bearing the mutation recovered the correct full-length SPINK5 m RNA and the corresponding functional lympho-epithelial Kazal-type related inhibitor protein in a dose-dependent manner. This study documents the reliability of a mutation-specific, Ex Spe U1-based, splicing therapy for a relatively large subset of European NS patients. Usage of Ex Spe U1 may represent a general approach for correction of splicing defects affecting skin disease genes. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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