Method of making transgenic mice lacking endogenous heavy chains
First Claim
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1. A method for producing a transgenic mouse lacking expression of an endogenous immunoglobulin heavy chain, comprising:
- (a) deleting the J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; and
(b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker,wherein the transgenic mouse lacks expression of an endogenous immunoglobulin heavy chain.
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Abstract
The subject invention provides non-human mammalian hosts characterized by inactivated endogenous Ig loci. The hosts are produced by repetitive transformations of embryonic stem cells by homologous recombination, preferably in conjunction with breeding. Different strategies are employed for recombination of the human loci randomly or at analogous host loci.
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Citations
13 Claims
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1. A method for producing a transgenic mouse lacking expression of an endogenous immunoglobulin heavy chain, comprising:
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(a) deleting the J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; and (b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker, wherein the transgenic mouse lacks expression of an endogenous immunoglobulin heavy chain. - View Dependent Claims (5, 13)
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2. A method for producing a transgenic mouse lacking expression of an endogenous immunoglobulin heavy chain, comprising:
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(a) deleting the J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; and (b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain an immunoglobulin heavy chain locus in which all of the J segment genes are deleted, wherein the transgenic mouse lacks expression of an endogenous immunoglobulin heavy chain.
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3. A method for producing a transgenic mouse and its progeny lacking expression of endogenous immunoglobulin heavy chains, comprising;
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(a) deleting the J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; (b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain an immunolobulin heavy chain locus in which all of the J segment genes are deleted, and (c) breeding the transgenic mouse as needed to produce a transgenic mouse and its progeny lacking expression of endogenous immunoglobulin heavy chains.
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4. A method for producing a transgenic mouse and its progeny lacking expression of endogenous immunoglobulin heavy chains, comprising:
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(a) deleting the J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; (b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker, and (c) breeding the transgenic mouse as needed to produce a transgenic mouse and its progeny lacking expression of endogenous immunoglobulin heavy chains.
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6. A method for preventing the expression of an endogenous immunoglobulin heavy chain in a transgenic mouse, comprising:
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(a) deleting the J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; and (b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker, wherein the transgenic mouse lacks expression of an endogenous immunoglobulin heavy chain. - View Dependent Claims (10)
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7. A method for preventing the expression of an endogenous immunoglobulin heavy chain in a transgenic mouse, comprising:
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(a) deleting the J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; and (b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain an endogenous immunoglobulin heavy chain locus in which all of the J segment genes are deleted, wherein the transgenic mouse lacks expression of an endogenous immunoglobulin heavy chain.
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8. A method for preventing the expression of endogenous immunoglobulin heavy chains in a transgenic mouse and its progeny, comprising:
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(a) deleting J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; (b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain an immunoglobulin heavy chain locus in which all of the J segment genes are deleted, and (c) breeding the transgenic mouse as needed to produce a transgenic mouse and its progeny lacking expression of endogenous immunoglobulin heavy chains.
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9. A method for preventing the expression of endogenous immunoglobulin heavy chains in a transgenic mouse and its progeny, comprising:
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(a) deleting J segment genes from at least one endogenous heavy chain locus in a mouse embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker; (b) producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker, and (c) breeding the transgenic mouse as needed to produce a transgenic mouse and its progeny lacking expression of endogenous immunoglobulin heavy chains.
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- 11. A method for producing a mouse embryonic stem cell with at least one inactivated endogenous immunoglobulin heavy chain locus, comprising deleting the J segment genes of at least one endogenous immunoglobulin heavy chain locus in the mouse embryonic stem cell to prevent rearrangement of the locus and to prevent the formation of a transcript of a rearranged immunoglobulin heavy chain, the deletion being effected by a targeting vector comprising a gene encoding a selectable marker.
Specification