Methods of generating knock-out rodents
OwnerMichael N. Gouldinventor
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Abstract
A method for generating knock-out rodents including rats and mice is disclosed. The method involves mutagenizing a rodent with a mutagen, obtaining progeny of the mutagenized rodent, and identifying, among the progeny, one progeny that carries a loss-of-function modification of a target gene. The preferred mutagen for generating knock-out mice and rats is N-ethyl-N-nitrosourea (ENU). The preferred screening assays for identifying a progeny of a mutagenized animal that carries a loss-of-function modification are yeast truncation assays and yeast functional assays. Knock-out rodents generated by the method of the present invention are also within the scope of the invention.
Claims36
A method for producing a knock-out rodent comprising the steps of:
- mutagenizing a rodent with a mutagen;
- obtaining progeny of the mutagenized rodent;
- and
- identifying, among the progeny, one progeny that carries a loss-of-function modification of a target gene.
A method for producing a knock-out rodent comprising the steps of:
- mutagenizing a rodent with a mutagen;
- obtaining progeny of the mutagenized rodent;
- collecting germ cells and optionally one or more other tissues from the progeny;
- identifying germ cells of a progeny that carry a loss-of-function modification of a target gene by analyzing a polynucleotide sample prepared from the germ cells or other tissues of the same progeny;
- and
- recovering a knock-out rodent from the germ cells identified in the identification step.
A knock-out rat comprising a loss of function modification of a pre-selected target gene in all of its germ cells and somatic cells wherein the loss of function modification is introduced into said rat, or an ancestor of said rat through genome manipulation.
A method for producing a knock-out mouse comprising the steps of:
- mutagenizing mouse embryonic stem cells with a mutagen;
- screening the embryonic stem cells by a biological screening method to identify one or more cells that carry a loss-of-function modification of a target gene;
- and
- recovering a knock-out mouse from a cell that carries a loss-of-function modification of a target gene identified in the screening step.
Dependencies are inferred from the claim text.