title: A novel technique based on a PNA hybridization probe and FRET principle for quantification of mutant genotype in fibrous dysplasia/McCune-Albright syndrome.
abstract: Somatic mutations are present in various proportions in numerous developmental pathologies. Somatic activating missense mutations of the GNAS gene encoding the Gs(alpha) protein have previously been shown to be the cause of fibrous dysplasia of bone (FD)/McCune-Albright syndrome (MAS). Because in MAS patients, tissues as diverse as melanocytes, gonads and bone are affected, it is generally accepted that the GNAS mutation in this disease must have occurred early in development. Interestingly, it has been shown that the development of an active FD lesion may require both normal and mutant cells. Studies of the somatic mosaic states of FD/MAS and many other somatic diseases need an accurate method to determine the ratio of mutant to normal cells in a given tissue. A new method for quantification of the mutant:normal ratio of cells using a PNA hybridization probe-based FRET technique was developed. This novel technique, with a linear sensitivity of 2.5% mutant alleles, was used to detect the percentage mutant cells in a number of tissue and cell culture samples derived from FD/MAS lesions and could easily be adapted for the quantification of mutations in a large spectrum of diseases including cancer.
authors.affiliation: Craniofacial and Skeletal Diseases Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892-4320, USA. akaradag@dir.nidcr.nih.gov
authors.forename: Abdullah
authors.initials: A
authors.lastname: Karadag
authors.forename: Mara
authors.initials: M
authors.lastname: Riminucci
authors.forename: Paolo
authors.initials: P
authors.lastname: Bianco
authors.forename: Natasha
authors.initials: N
authors.lastname: Cherman
authors.forename: Sergei A
authors.initials: SA
authors.lastname: Kuznetsov
authors.forename: Nga
authors.initials: N
authors.lastname: Nguyen
authors.forename: Michael T
authors.initials: MT
authors.lastname: Collins
authors.forename: Pamela G
authors.initials: PG
authors.lastname: Robey
authors.forename: Larry W
authors.initials: LW
authors.lastname: Fisher
mesh_terms: D000483:Alleles; D001483:Base Sequence; D001842:Bone and Bones / Q000473:pathology; D002864:Chromogranins; D004252:DNA Mutational Analysis / Q000379:methods*; D015342:DNA Probes / Q000235:genetics*; D005357:Fibrous Dysplasia of Bone / Q000235:genetics*; D005359:Fibrous Dysplasia, Polyostotic / Q000235:genetics*; D031541:Fluorescence Resonance Energy Transfer; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000235:genetics; D005838:Genotype; D006801:Humans; D009154:Mutation / Q000235:genetics*; D020135:Peptide Nucleic Acids / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012015:Reference Standards; D012680:Sensitivity and Specificity
chemical_list: D002864:Chromogranins; D015342:DNA Probes; D020135:Peptide Nucleic Acids; C406732:GNAS protein, human; D019205:GTP-Binding Protein alpha Subunits, Gs