Method for producing genome-edited cells utilizing homologous recombination
AssigneeOsaka University
InventorShinichiro NAKADA
Life of this patent
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Abstract
A genome editing method utilizing recombination between homologous chromosomes having different bases at the target site is disclosed. Specifically, (1) a method of using cells in which the function of a gene selected from the group of Lig4 gene, PARP1 gene, XRCC1 gene, MSH2 gene, and SMARCAL1 gene is suppressed, or (2) a method of causing a single-strand break at one site in the neighboring DNA region of the target site on the chromosome of the recipient, and causing a single-strand break at one site different from the site corresponding to the site where the single-strand break is caused on the chromosome of the recipient, on the chromosome of the donor.
Claims8
A method for producing genome-edited cells, comprising:
- introducing a combination of site-specific nickases that causes single-strand breaks in a neighboring DNA region of a specific site in homologous chromosomes having different bases between the homologous chromosomes at the specific site, inducing homologous recombination using one of the homologous chromosomes as a recipient and the other as a donor, and substituting a base of the recipient with a base of the donor at the specific site, wherein
- the cells have suppressed function of a gene selected from the group consisting of Lig4 gene, PARP1 gene, XRCC1 gene, MSH2 gene, and SMARCAL1 gene, and
- the combination of site-specific nickases causes single-strand breaks at multiple sites in the neighboring DNA region of the specific site on the chromosome of the recipient and causes a single-strand break at one site corresponding to the site where the single-strand breaks are caused on the chromosome of the recipient, on the chromosome of the donor.
A method for producing genome-edited cells, comprising:
- introducing a combination of site-specific nickases that causes single-strand breaks in a neighboring DNA region of a specific site in homologous chromosomes having different bases between the homologous chromosomes at the specific site, inducing homologous recombination using one of the homologous chromosomes as a recipient and the other as a donor, and substituting a base of the recipient with a base of the donor at the specific site, wherein
- the combination of site-specific nickases causes a single-strand break at one site in the neighboring DNA region of the specific site on the chromosome of the recipient and causes a single-strand break at a different site from the site corresponding to the site where the single-strand break is caused on the chromosome of the recipient, at one site on the chromosome of the donor.
Dependencies are inferred from the claim text.