Non-human animals having a humanized signal-regulatory protein gene
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsAndrew J. Murphy, O. Gavin Thurston, Bindu Varghese, Cagan Gurer
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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.
Claims41
A rodent whose genome comprises a replacement of exons 2, 3 and 4 of a rodent SIRPα gene at an endogenous rodent 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 rodent SIRPα promoter at said endogenous rodent SIRPα locus, and expresses in said rodent 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 rodent SIRPα protein encoded by said rodent SIRPα gene, and wherein the rodent is selected from a mouse or a rat.
An isolated rodent cell or tissue whose genome comprises a replacement of exons 2, 3 and 4 of a rodent SIRPα gene at an endogenous rodent 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 rodent SIRPα promoter at said endogenous rodent SIRPα locus, and expresses in said rodent cell or tissue 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 rodent SIRPα protein encoded by said rodent SIRPα gene, and wherein the rodent is selected from a mouse or a rat.
A method of making a rodent, the method comprising:
- (a) replacing exons 2, 3 and 4 of a rodent SIRPα gene at an endogenous rodent SIRPα locus in a rodent 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 rodent SIRPα promoter at said endogenous rodent 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 rodent SIRPα protein encoded by said rodent SIRPα gene, and wherein the rodent is selected from mouse or rat, thereby obtaining a modified rodent ES cell comprising said humanized SIRPα gene;
- (b) creating a rodent using the modified ES cell of (a).
A method of engrafting human cells into a rodent, comprising steps of:
- (a) providing a rodent whose genome comprises a replacement of exons 2, 3 and 4 of a rodent SIRPα gene at an endogenous rodent 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 rodent SIRPα promoter at said endogenous rodent SIRPα locus and expresses in said rodent 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 rodent SIRPα protein encoded by said rodent SIRPα gene, and wherein the rodent is selected from a mouse or a rat;
- and
- (b) transplanting one or more human cells into the rodent.
A method comprising,
- (a) providing one or more rodent cells whose genome comprises a replacement of exons 2, 3 and 4 of a rodent SIRPα gene at an endogenous rodent 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 rodent SIRPα promoter at said endogenous rodent SIRPα locus and expresses in said one or more rodent 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 rodent SIRPα protein encoded by said rodent SIRPα gene, and wherein the rodent is selected from a mouse or a rat;
- (b) incubating the one or more rodent cells of step (a) with a labeled substrate;
- and
- (c) measuring phagocytosis of the labeled substrate by the one or more rodent cells of step (b).
A method comprising,
- (a) providing a rodent whose genome comprises a replacement of exons 2, 3 and 4 of a rodent SIRPα gene at an endogenous rodent 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 rodent SIRPα promoter at said endogenous rodent SIRPα locus and expresses in said rodent 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 rodent SIRPα protein encoded by said rodent SIRPα gene, and wherein the rodent is selected from a mouse or a rat;
- (b) exposing the rodent to an antigen;
- and
- (c) measuring phagocytosis of the antigen by one or more cells of the rodent.
A method of assessing the therapeutic efficacy of a drug targeting human cells, comprising:
- providing a rodent whose genome a replacement of exons 2, 3 and 4 of a rodent SIRPα gene at an endogenous rodent 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 rodent SIRPα promoter at said endogenous rodent SIRPα locus and expresses in said rodent 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 rodent SIRPα protein encoded by said rodent SIRPα gene, and wherein the rodent is selected from a mouse or a rat;
- transplanting one or more human cells into the rodent;
- administering a drug candidate to said rodent;
- and
- monitoring the human cells in the rodent to determine the therapeutic efficacy of the drug candidate.
Dependencies are inferred from the claim text.