Simple method for detcting pluripotent stem cells genetically modified by homologous recombination
AssigneeKyoto University
InventorsShinya Yamanaka, Kenji Osafune
Life of this patent
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Abstract
The present invention relates to a method for producing pluripotent stem cells, which comprises the steps of introducing an artificial chromosome having a genetically modified chromosome fragment as a targeting vector, and determining the number of some or all copies of the introduced artificial chromosome using an SNP array, so as to select pluripotent stem cells modified by homologous recombination, and, a method for detecting pluripotent stem cells genetically modified by homologous recombination.
Claims13
A method for producing pluripotent stem cells genetically modified by homologous recombination, comprising the following steps (1) to (3) of:
- (1) introducing an artificial chromosome having a genetically modified chromosome fragment into pluripotent stem cells, so as to prepare a population consisting of pluripotent stem cell clones assumed to be genetically modified;
- (2) determining the number of some or all copies of the introduced artificial chromosome using an SNP array for the population of pluripotent stem cell clones;
- and
- (3) selecting pluripotent stem cell clones in which the number of copies is equivalent to or lower than the same number of wild-type cells into which no artificial chromosomes have been introduced, as pluripotent stem cells genetically modified by homologous recombination.
A method for detecting pluripotent stem cells genetically modified by homologous recombination, comprising a step of determining, when pluripotent stem cells genetically modified by homologous recombination are produced, the number of copies of a recombined region using an SNP array, and then detecting pluripotent stem cell clones in which the number of copies is equivalent to or lower than the same number of wild-type cells not modified by homologous recombination, as pluripotent stem cells genetically modified by homologous recombination.
Dependencies are inferred from the claim text.