Abstract |
An intronic point mutation was identified in the ocular albinism type 1 (OA1) gene (HUGO symbol, GPR143) in a family with the X-linked form of ocular albinism. Interestingly, the mutation creates a new acceptor splice site in intron 7 of the OA1 gene. In addition to low levels of normally spliced mRNA product of the OA1 gene, the patient samples contained also an aberrantly spliced mRNA with a 165 bp fragment of intron 7 (from position +750 to +914) inserted between exons 7 and 8. The abnormal transcript contained a premature stop codon and was unstable, as revealed by Northern blot analysis. We defined that mutation NC_000023.8:g.25288G>A generated a consensus binding motif for the splicing factor enhancer ASF/SF2, which most likely favored transcription of the aberrant mRNA. Furthermore, it activated a cryptic donor-splice site causing the inclusion between exons 7 and 8 of the 165 bp intronic fragment. Thus, the aberrant splicing is most likely explained by the generation of a de novo splicing enhancer motif. Finally, to rescue OA1 expression in the patient's melanocytes, we designed an antisense morpholino modified oligonucleotide complementary to the mutant sequence. The morpholino oligonucleotide (MO) was able to rescue OA1 expression and restore the OA1 protein level in the patient's melanocytes through skipping of the aberrant inclusion. The use of MO demonstrated that the lack of OA1 was caused by the generation of a new splice site. Furthermore, this technique will lead to new approaches to correct splice site mutations that cause human diseases.
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Authors | Francesco Vetrini, Roberta Tammaro, Sergio Bondanza, Enrico M Surace, Alberto Auricchio, Michele De Luca, Andrea Ballabio, Valeria Marigo |
Journal | Human mutation
(Hum Mutat)
Vol. 27
Issue 5
Pg. 420-6
(May 2006)
ISSN: 1098-1004 [Electronic] United States |
PMID | 16550551
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
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Copyright | Published 2006 Wiley-Liss, Inc. |
Chemical References |
- Eye Proteins
- GPR143 protein, human
- Membrane Glycoproteins
- Morpholines
- Oligonucleotides, Antisense
- RNA Splice Sites
- RNA, Messenger
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Topics |
- Albinism, Ocular
(genetics)
- Base Sequence
- DNA Mutational Analysis
- Eye Proteins
(drug effects, genetics)
- Humans
- Melanocytes
(cytology, drug effects, metabolism)
- Membrane Glycoproteins
(drug effects, genetics)
- Molecular Sequence Data
- Morpholines
(chemistry)
- Oligonucleotides, Antisense
(pharmacology)
- Point Mutation
- RNA Splice Sites
(genetics)
- RNA Splicing
- RNA, Messenger
(drug effects, metabolism)
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