Mono-allelic mutation analysis for identifying germline mutations
AssigneeThe Johns Hopkins University
InventorsBert Vogelstein, Kenneth W. Kinzler, Nickolas Papadopoulos
Life of this patent
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Abstract
A diagnostic strategy for detection of inherited diseases caused by germline mutations is based on somatic cell hybridization. Each allele of a human gene involved in the inherited disease is isolated in a somatic cell hybrid. The products of the isolated human allele are then observed in the absence of the other allele of the human.
Claims97
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- obtaining cells of the human;
- fusing said cells to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome, to form human-rodent cell hybrids;
- testing said human-rodent cell hybrids to confirm the presence of said chromosome of the human in said hybrid;
- testing said hybrids which contains contain said chromosome to detect a protein product of said first gene, absence of said protein product or diminished amounts of said protein product indicating the presence of a mutation in the first gene of the human.
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- obtaining peripheral blood lymphocytes of the human;
- fusing said peripheral blood lymphocytes to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome, to form human-rodent cell hybrids;
- testing said human-rodent cell hybrids to confirm the presence of said chromosome of the human in said hybrids, wherein the presence of said chromosome is tested by identifying the presence of microsatellite markers flanking the first gene;
- testing said hybrids which contains contain said chromosome to detect a protein product of said first gene by Western blotting, absence of said protein product or diminished amounts of said protein product indicating the presence of a mutation in the first gene of the human.
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- fusing cells of the human to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome, to form human-rodent cell hybrids;
- testing said human-rodent cell hybrids to confirm the presence of said chromosome of the human in said hybrid;
- testing said hybrids which contain said chromosome to detect a protein product of said first gene, absence of said protein product or diminished amounts of said protein product indicating the presence of a mutation in the first gene of the human.
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- (a) fusing cells of the human to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome, to form human-rodent cell hybrids;
- (b) testing said human-rodent cell hybrids to confirm the presence of said chromosome of the human in said hybrid;
- (c) testing said hybrids formed in step (a) which contain said chromosome to detect a mutation in the first gene of the human.
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- fusing cells of the human to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome, to form human-rodent cell hybrids;
- testing said human-rodent cell hybrids to confirm the presence of said chromosome of the human in said hybrid;
- testing said hybrids which contain said chromosome to detect a haplotype, wherein the haplotype has been associated with a mutation in the first gene.
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- testing human-rodent cell hybrids formed by:
- fusing cells of the human to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome,
- to confirm the presence of said chromosome of the human in said hybrid;
- testing said human-rodent cell hybrids which contain said chromosome to detect a protein product of said first gene, absence of said protein product or diminished amounts of said protein product indicating the presence of a mutation in the first gene of the human.
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- testing human-rodent cell hybrids formed by:
- fusing cells of the human to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome,
- to confirm the presence of said chromosome of the human in said hybrid, wherein the presence of said chromosome is tested by identifying the presence of microsatellite markers flanking the first gene;
- testing said human-rodent cell hybrids which contain said chromosome to detect a protein product of said first gene by Western blotting, absence of said protein product or diminished amounts of said protein product indicating the presence of a mutation in the first gene of the human.
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- (a) testing human-rodent cell hybrids formed by:
- fusing cells of the human to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome,
- to confirm the presence of said chromosome of the human in said hybrid;
- (b) testing said hybrids formed in step (a) which contain said chromosome to detect a mutation in the first gene of the human.
A method of detecting mutations in a first gene of a human, wherein the first gene has been previously mapped to a human chromosome, comprising the steps of:
- testing human-rodent cell hybrids formed by:
- fusing cells of the human to rodent cell recipients which carry a mutation which can be complemented by a second gene which has been mapped to said chromosome,
- to confirm the presence of said chromosome of the human in said hybrid;
- testing said hybrids which contain said chromosome to detect a haplotype, wherein the haplotype has been associated with a mutation in the first gene.
Dependencies are inferred from the claim text.