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US 2002/0119439 A1Published application
Genetic assays for trinucleotide repeat mutations in eukaryotic cells
AssigneeBoard of Regents of the University of Nebraska
InventorsRobert S. Lahue, Juan J. Miret, Richard Pelletier
Life of this patent
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Abstract
A method for assaying trinucleotide repeat mutations in eukaryotic cells is provided.
Claims22
A method for detecting alterations in trinucleotide repeat tract lengths, comprising the steps of:
- a) contacting mammalian cells with a shuttle vector under conditions whereby said shuttle vector enters and replicates in said cells;
- b) recovering said replicated shuttle vector from said mammalian cells;
- c) introducing said recovered shuttle vector into a yeast cell in the presence of a selection agent, alteration of said trinucleotide repeat tract conferring resistance to said selective agent;
- and
- d) selecting yeast cells comprising said shuttle vector containing trinucleotide repeat tract alterations which survive the presence of said selection agent.
2 more elements
A method for identifying trinucleotide repeat tract expansions, comprising the steps of:
- a) contacting mammalian cells with a shuttle vector containing trinucleotide repeat tract length of approximately 25 repeats under conditions whereby said shuttle vector enters and replicates in said cells;
- b) recovering said replicated shuttle vector from said mammalian cells;
- c) introducing said recovered shuttle vector into a yeast cell in the absence of histidine, expansion of said trinucleotide repeat tract conferring a His+ phenotype;
- and
- d) selecting yeast cells comprising said shuttle vector containing trinucleotide repeat tract expansions which survive in the absence of histidine.
2 more elements
A method for identifying contractions in trinucleotide repeat tract lengths, comprising the steps of:
- a) contacting mammalian cells with a shuttle vector containing trinucleotide repeat tract length of approximately 33 repeats under conditions whereby said shuttle vector enters and replicates in said cells;
- b) recovering said replicated shuttle vector from said mammalian cells;
- c) introducing said recovered shuttle vector into a yeast cell in the absence of uracil, expansion of said trinucleotide repeat tract conferring a Ura+ phenotype;
- and
- d) selecting yeast cells comprising said shuttle vector containing trinucleotide repeat tract contractions which survive in the absence of uracil.
2 more elements
A method for identifying contractions in trinucleotide repeat tract lengths, comprising the steps of:
- a) contacting mammalian cells with a shuttle vector containing trinucleotide repeat tract length of approximately 50 repeats under conditions whereby said shuttle vector enters and replicates in said cells;
- b) recovering said replicated shuttle vector from said mammalian cells;
- c) introducing said recovered shuttle vector into a yeast cell in the absence of uracil, expansion of said trinucleotide repeat tract conferring a Ura+ phenotype;
- and
- d) selecting yeast cells comprising said shuttle vector containing trinucleotide repeat tract contractions which survive in the absence of uracil.
2 more elements
Dependencies are inferred from the claim text.