Directed genetic modifications of human stem cells
First Claim
1. A method of performing targeted modifications of human embryonic stem (ES) cells, the method comprising the steps of obtaining copies of a genetic construct which includes a foreign gene and which has regions at either of its ends homologous with a pair of selected regions in the genome of the ES cells so that homologous recombination will occur between the genetic construct and the selected regions of the genome of the stem cells;
- electroporating the copies of the genetic construct into stem cell in culture; and
identifying cells which contain the genetic construct.
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Accused Products
Abstract
Human embryonic stem cells can be genetically transformed by a combination of electroporation and homologous recombination. This technique makes it possible to create targeted inserts or deletions to the genome of the stem cells. This ability makes it possible to create populations of progeny cells which have differentiated into a target cell type of a specific desired lineage.
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Citations
16 Claims
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1. A method of performing targeted modifications of human embryonic stem (ES) cells, the method comprising the steps of
obtaining copies of a genetic construct which includes a foreign gene and which has regions at either of its ends homologous with a pair of selected regions in the genome of the ES cells so that homologous recombination will occur between the genetic construct and the selected regions of the genome of the stem cells; -
electroporating the copies of the genetic construct into stem cell in culture; and
identifying cells which contain the genetic construct. - View Dependent Claims (2, 3, 4)
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5. Human cells in culture derived from human embryonic stem cells, the cells comprising in their genome an inserted genetic construct which knocks out the functioning of a gene which would otherwise be expressed in those human cells in culture.
- 6. Human cells in culture derived from human embryonic stem cells, the cells comprising in their genome an inserted genetic construct which introduced a mutation into a native gene in those human cells in culture.
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7. A method of purifying cells of a defined lineage from cultures of human embryonic stem (ES) cells, the method comprising the steps of
obtaining copies of a genetic construct which has regions at either of its ends homologous with a pair of selected regions in the genome of the ES cells so that homologous recombination will occur between the genetic construct and the selected regions of the genome of the stem cells, the genetic construct including a marker gene which will be expressed only in cells of the defined lineage; -
electroporating the copies of the genetic construct into stem cell in culture; and
identifying cells which express the marker gene in the genetic construct and purifying those cells from other cells. - View Dependent Claims (8, 9, 10, 11)
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- 14. Human cells in culture derived from human embryonic stem cells, the cells comprising in their genome an inserted genetic construct which expresses an inserted gene only when the human cells are in a desired state of differentiation.
Specification