GM25541
iPSC from Fibroblast
Description:
LESCH-NYHAN SYNDROME; LNS
HYPOXANTHINE GUANINE PHOSPHORIBOSYLTRANSFERASE 1; HPRT1; HPRT
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Repository
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NIGMS Human Genetic Cell Repository
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| Subcollection |
Heritable Diseases |
| Protocols |
Protocol PDF |
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Biopsy Source
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Skin
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Cell Type
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Stem cell
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Cell Subtype
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Induced pluripotent stem cell
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Transformant
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Reprogrammed (Episomal)
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Sample Source
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iPSC from Fibroblast
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Race
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White
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Family Member
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1
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Relation to Proband
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proband
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Confirmation
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Clinical summary/Case history
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ISCN
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46,XY.arr[hg19](1-22)x2,(XY)x1
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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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| Passage Frozen |
14 |
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| Induced Pluripotent Stem Cell |
The frozen cell line submitted to the Repository was recovered and expanded. The expanded line was evaluated for viability surface antigen expression and alkaline phosphatase activity. Pluripotency was assessed via embryoid body (EB) formation. Steady-state mRNA expression patterns of undifferentiated iPSC and EBs were determined via real-time PCR. Characterization data are included in the Certificate of Analysis. |
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| Gene |
HPRT1 |
| Chromosomal Location |
Xq26-q27.2 |
| Allelic Variant 1 |
308000.0047; LESCH-NYHAN SYNDROME |
| Identified Mutation |
EX2-3DUP, IVS1DEL; In GM1662 and GM6804 from patients with LNS, Yang et al.(1984, 1988) found a complex rearrangement involving duplication of exons 2 and 3 and deletion of intron 1. Increased size of mRNA was observed. Monnat et al. (1992) demonstrated that the duplication in GM6804 was generated by the nonhomologous insertion of duplicated HPRT DNA into HPRT intron 1. They found that the duplication was genetically unstable and had a reversion rate approximately 100-fold higher than the rate of duplication formation. Exons 2 and 3, together with 13.7 kb of surrounding HPRT sequence, were duplicated.
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| Remarks |
iPSC line derived from GM01662 Fibroblast by episomal reprogramming; Clinically affected; no detectable HPRT and normal APRT activity; has an exon 2,3 duplication in the HPRT1 gene and a correspondingly elongated HPRT mRNA; atypical disease: lacks compulsive self-mutilation and mental retardation; see GM06804 for Lymph of same subject; mother (GM01659) and sisters (GM01660/GM01661) are unaffected carriers. Genomic integration of episomal plasmid detected; however there was no expression of exogenous reprogramming factors. Researchers purchasing hiPSCs from the NIGMS Repository are responsible for any limited use label licenses (LULLs) applicable to the cell line purchased. The applicable LULL to this line is iPS Academia Japan, Inc.. |
| Shammas MA, Koley H, Beer DG, Li C, Goyal RK, Munshi NC, Growth arrest, apoptosis, and telomere shortening of Barrett's-associated adenocarcinoma cells by a telomerase inhibitor. Gastroenterology126(5):1337-46 2004 |
| PubMed ID: 15131795 |
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| Shammas MA, Shmookler Reis RJ, Akiyama M, Koley H, Chauhan D, Hideshima T, Goyal RK, Hurley LH, Anderson KC, Munshi NC, Telomerase inhibition and cell growth arrest by G-quadruplex interactive agent in multiple myeloma. Mol Cancer Ther2(9):825-33 2003 |
| PubMed ID: 14555701 |
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| Hotamisligil GS, Breakefield XO, Human monoamine oxidase A gene determines levels of enzyme activity. Am J Hum Genet49:383-92 1991 |
| PubMed ID: 1678250 |
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| Yang TP, Stout JT, Konecki DS, Patel PI, Alford RL, Caskey CT, Spontaneous reversion of novel Lesch-Nyhan mutation by HPRT gene rearrangement. Somat Cell Mol Genet14:293-303 1988 |
| PubMed ID: 2835825 |
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| Caskey CT, Disease diagnosis by recombinant DNA methods. Science236:1223-9 1987 |
| PubMed ID: 3296189 |
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| Gunby JL, Tomkins DJ, Chang PL, Somatic cell hybridization of Roberts syndrome and normal human fibroblasts transfected with plasmids carrying dominant selection markers. Somat Cell Mol Genet13:245-52 1987 |
| PubMed ID: 3474785 |
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| Chang PL, Gunby JL, Tomkins DJ, Mak I, Rosa NE, Mak S, Transformation of human cultured fibroblasts with plasmids carrying dominant selection markers and immortalizing potential. Exp Cell Res167:407-16 1986 |
| PubMed ID: 3021482 |
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| Boggs BA, Nussbaum RL, Two anonymous X-specific human sequences detecting restriction fragment length polymorphisms in region Xq26----qter. Somat Cell Mol Genet10:607-13 1984 |
| PubMed ID: 6095463 |
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| Drescher-Lincoln CK, Smith JR, Inhibition of DNA synthesis in senescent-proliferating human cybrids is mediated by endogenous proteins. Exp Cell Res153:208-17 1984 |
| PubMed ID: 6734738 |
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| Yang TP, Patel PI, Chinault AC, Stout JT, Jackson LG, Hildebrand BM, Caskey CT, Molecular evidence for new mutation at the hprt locus in Lesch-Nyhan patients. Nature310:412-4 1984 |
| PubMed ID: 6087154 |
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| Nussbaum RL, Crowder WE, Nyhan WL, Caskey CT, A three-allele restriction-fragment-length polymorphism at the hypoxanthine phosphoribosyltransferase locus in man. Proc Natl Acad Sci U S A80:4035-9 1983 |
| PubMed ID: 6306659 |
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| Pereira-Smith OM, Smith JR, Evidence for the recessive nature of cellular immortality. Science221:964-6 1983 |
| PubMed ID: 6879195 |
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| Pereira-Smith OM, Smith JR, Phenotype of low proliferative potential is dominant in hybrids of normal human fibroblasts. Somatic Cell Genet8:731-42 1982 |
| PubMed ID: 7163953 |
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| James L, Veomett GE, Decreased longevity of human diploid cells after incorporation of latex spheres within their cytoplasm. Exp Cell Res132:468-73 1981 |
| PubMed ID: 7215456 |
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| Pereira-Smith OM, Smith JR, Expression of SV40 T antigen in finite life-span hybrids of normal and SV40-transformed fibroblasts. Somatic Cell Genet7:411-21 1981 |
| PubMed ID: 6269237 |
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| Schonberg S, German J, Sister chromatid exchange in cells metabolically coupled to Bloom's syndrome cells. Nature284:72-4 1980 |
| PubMed ID: 7354874 |
| Passage Frozen |
14 |
| Split Ratio |
1:6 |
| Temperature |
37 C |
| Percent CO2 |
5% |
| Percent O2 |
AMBIENT |
| Medium |
Ham's F12 Medium/Dulbecco Modified Eagles Medium, 1:1 mixture with 2mM L-glutamine or equivalent |
| Serum |
20% Knock-out Serum Replacement Not inactivated |
| Substrate |
Gelatin + Feeder Layer |
| Supplement |
Basic Fibroblast Growth Factor 10 ng/ml |
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