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Confirmation of homozygosity for a single nucleotide substitution mutation in a Cockayne syndrome patient using monoallelic mutation analysis in somatic cell hybrids
Author(s) -
McDaniel Lisa D.,
Legerski Randy,
Lehmann Alan R.,
Friedberg Errol C.,
Schultz Roger A.
Publication year - 1997
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/(sici)1098-1004(1997)10:4<317::aid-humu8>3.0.co;2-d
Subject(s) - genetics , biology , mutation , disease gene identification , allele , germline mosaicism , complementation , gene , exome sequencing , phenotype
The identification of individuals homozygous for a specific mutation offers advantages for the elucidation of molecular mechanisms of hereditary disease states. Cockayne syndrome is a rare autosomal recessive disorder, the molecular basis of which is complicated by significant genetic and clinical heterogeneity. The genes associated with both genetic complementation groups, CSA and CS‐B, have been identified. We have previously identified a number of CSA mutations, including a single base substitution that introduces a stop codon ( 322 Tyr→Stop) mutation in the C‐terminal region for at least one allele of the CSA gene in a severely affected patient. We now present data confirming the existence of homozygosity in this patient using a strategy with general applicability. Somatic cell hybrids were established by fusing patient cells with mouse A9 cells. Screening with chromosome 5 specific polymorphic markers facilitated identification of hybrid clones bearing only one of the distinct CSA alleles. Sequencing of a portion of the human CSA gene in a subset of these hybrids permitted monoallelic mutation analysis and confirmed the presence of the 322 Tyr→Stop mutation in both alleles. Hum Mutat 10:317–321, 1997. © 1997 Wiley‐Liss, Inc.