Back to results
US 9,193,977 B2Granted
Non-human animals having a humanized signal-regulatory protein gene
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsAndrew J. Murphy, O. Gavin Thurston, Bindu Varghese, Cagan Gurer
Life of this patent
Estimating the term…
Abstract
Genetically modified non-human animals and methods and compositions for making and using the same are provided, wherein the genetic modification comprises a humanization of an endogenous signal-regulatory protein gene, in particular a humanization of a SIRPα gene. Genetically modified mice are described, including mice that express a human or humanized SIRPα protein from an endogenous SIRPα locus.
Claims28
A method of making a mouse, the method comprising:
- (a) replacing exons 2, 3 and 4 of a mouse SIRPα gene at an endogenous mouse SIRPα locus in a mouse ES cell with exons 2, 3 and 4 of a human SIRPα gene to form a humanized SIRPα gene, wherein said humanized SIRPα gene is operably linked to a mouse SIRPα promoter at said endogenous mouse SIRPα locus and encodes a humanized SIRPα protein comprising an extracellular portion of the human SIRPα protein encoded by said human SIRPα gene and an intracellular portion of the mouse SIRPα protein encoded by said mouse SIRPα gene, thereby obtaining a modified mouse ES cell comprising said humanized SIRPα gene;
- (b) creating a mouse using the modified ES cell of (a).
A method of engrafting human cells into a mouse, comprising steps of:
- (a) providing a mouse whose genome comprises a replacement of exons 2, 3 and 4 of a mouse SIRPα gene at an endogenous mouse SIRPα locus with exons 2, 3 and 4 of a human SIRPα gene to form a humanized SIRPα gene, wherein said humanized SIRPα gene is operably linked to a mouse SIRPα promoter at said endogenous mouse SIRPα locus and expresses in said mouse a humanized SIRPα protein comprising an extracellular portion of the human SIRPα protein encoded by said human SIRPα gene and an intracellular portion of the mouse SIRPα protein encoded by said mouse SIRPα gene;
- and
- (b) transplanting one or more human cells into the mouse.
A method comprising,
- (a) providing one or more mouse cells whose genome comprises a replacement of exons 2, 3 and 4 of a mouse SIRPα gene at an endogenous mouse SIRPα locus with exons 2, 3 and 4 of a human SIRPα gene to form a humanized SIRPα gene, wherein said humanized SIRPα gene is operably linked to a mouse SIRPα promoter at said endogenous mouse SIRPα locus and expresses in said one or more mouse cells a humanized SIRPα protein comprising an extracellular portion of the human SIRPα protein encoded by said human SIRPα gene and an intracellular portion of the mouse SIRPα protein encoded by said mouse SIRPα gene;
- (b) incubating the one or more mouse cells of step (a) with a labeled substrate;
- and
- (c) measuring phagocytosis of the labeled substrate by the one or more mouse cells of step (b).
1 more element
A method comprising,
- (a) providing a mouse whose genome comprises a replacement of exons 2, 3 and 4 of a mouse SIRPα gene at an endogenous mouse SIRPα locus with exons 2, 3 and 4 of a human SIRPα gene to form a humanized SIRPα gene, wherein said humanized SIRPα gene is operably linked to a mouse SIRPα promoter at said endogenous mouse SIRPα locus and expresses in said mouse a humanized SIRPα protein comprising an extracellular portion of the human SIRPα protein encoded by said human SIRPα gene and an intracellular portion of the mouse SIRPα protein encoded by said mouse SIRPα gene;
- (b) exposing the mouse to an antigen;
- and
- (c) measuring phagocytosis of the antigen by one or more cells of the mouse.
1 more element
A method of assessing the therapeutic efficacy of a drug targeting human cells, comprising:
- providing a mouse whose genome comprises a replacement of exons 2, 3 and 4 of a mouse SIRPα gene at an endogenous mouse SIRPα locus with exons 2, 3 and 4 of a human SIRPα gene to form a humanized SIRPα gene, wherein said humanized SIRPα gene is operably linked to a mouse SIRPα promoter at said endogenous mouse SIRPα locus and expresses in said mouse a humanized SIRPα protein comprising an extracellular portion of the human SIRPα protein encoded by said human SIRPα gene and an intracellular portion of the mouse SIRPα protein encoded by said mouse SIRPα gene;
- transplanting one or more human cells into the mouse;
- administering a drug candidate to said mouse;
- and
- monitoring the human cells in the mouse to determine the therapeutic efficacy of the drug candidate.
2 more elements
Dependencies are inferred from the claim text.