GM21941
Fibroblast from Skin, Unspecified
Description:
MARFAN SYNDROME; MFS
FIBRILLIN 1; FBN1
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Repository
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NIGMS Human Genetic Cell Repository
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| Subcollection |
Heritable Diseases |
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Biopsy Source
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Unspecified
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Cell Type
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Fibroblast
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Tissue Type
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Skin
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Transformant
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Untransformed
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Sample Source
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Fibroblast from Skin, Unspecified
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Relation to Proband
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proband
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Confirmation
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Molecular characterization before cell line submission to CCR
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Species
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Homo sapiens
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Common Name
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Human
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Remarks
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| PDL at Freeze |
7.35 |
| Passage Frozen |
7 |
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| IDENTIFICATION OF SPECIES OF ORIGIN |
Species of Origin confirmed by LINE assay |
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| Gene |
FBN1 |
| Chromosomal Location |
15q21.1 |
| Allelic Variant 1 |
134797.0030; MARFAN SYNDROME, CLASSIC |
| Identified Mutation |
6354C>T,EX51DEL,ILE2118ILE; Liu et al. (1997) carried out a systematic mutation search of PCR-amplified transcripts of the FBN1 gene from patients with Marfan syndrome. By long RT-PCR and restriction enzyme digestions, they identified skipping of FBN1 exons in 10% of Marfan cases. All but 1 of the these were due to sequence alterations at splice sites. In skin fibroblasts derived from a patient with classic Marfan syndrome, an abnormally migrating restriction fragment was identified and found to represent deletion of 66 bp due to in-frame skipping of the entire exon 51. This exon encodes the 3-prime portion of 1 of the 7 8-cysteine domains in FBN1 that is similar to a motif found in transforming growth factor-beta-1 binding protein (150390). Sequencing of exon 51 and surrounding splice sites, amplified from genomic DNA with intron primers, identified only 1 sequence variation unique to this sample: a C-to-T transition (6354C-T) at position +41 of exon 51. This mutation changes codon 2118 from AUC to AUU, both of which encode isoleucine. Liu et al. (1997) stated that this nucleotide change is unlikely to affect known binding sites of the splicing machinery. Further studies indicated that the skipping of exon 51 in these cells was due solely to the silent mutation, 6354C-T. Skipping of exon 51 associated with a 6339T-G mutation that changes a tyrosine (TAT) to a termination (TAG) codon (134797.0008) was previously reported as the cause of exon 51 skipping (Dietz et al., 1993; Dietz and Kendzior, 1994). Liu et al. (1997) commented that skipping caused by a silent mutation suggests the existence of an alternative mechanism of exon skipping yet to be discovered.
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| Remarks |
Clinically affected; classic features; donor subject has a C>T transition (silent mutation) at nucleotide 6354 in exon 51 of the FBN1 gene (6354C>T) resulting in the in-frame skipping of exon 51 (del 66 bp) |
| Liu W, Qian C, Francke U, Silent mutation induces exon skipping of fibrillin-1 gene in Marfan syndrome Nature genetics16:328-9 1997 |
| PubMed ID: 9241263 |
| Passage Frozen |
7 |
| Split Ratio |
1:4 |
| Temperature |
37 C |
| Percent CO2 |
10% |
| Percent O2 |
AMBIENT |
| Medium |
Dulbecco Modified Eagles Medium (high glucose) with 2mM L-glutamine or equivalent |
| Serum |
10% fetal bovine serum Not inactivated |
| Substrate |
None specified |
| Supplement |
- |
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