# US 10,477,842 B2 · Humanized IL-4 and IL-4Rα animals > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,477,842 B2 - Document ID: US10477842B2 - Status: Granted - Kind: B2 - Application number: 15/705,326 - Filed: 2017-09-15 - Published: 2018-01-04 - Granted: 2019-11-19 - Earliest claimed: 2014-05-07 - Assignees: Regeneron Pharmaceuticals, Inc. - Applicants: Regeneron Pharmaceuticals, Inc. - Inventors: Li-Hsien Wang; Yingzi Xue; Andrew J. Murphy; Sean Stevens - Web page: https://patentgrep.com/patent/US10477842B2 ## CPC - A01K 67/0278: Knock-in vertebrates, e.g. humanised vertebrates - C07K 14/5406: {IL-4} - C07K 14/7155: {for interleukins [IL]} - A01K 2207/15: Humanized animals - A01K 2217/072: maintaining or altering function, i.e. knock in - A01K 2217/20: Animal model comprising regulated expression system - A01K 2227/105: Murine - A01K 2267/01: Animal expressing industrially exogenous proteins - A01K 2267/0368: Animal model for inflammation - A01K 2267/0393: Animal model comprising a reporter system for screening tests - A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract Non-human animals comprising a human or humanized IL-4 and/or IL-4Rα nucleic acid sequence are provided. Non-human animals that comprise a replacement of the endogenous IL-4 gene and/or IL-4Rα gene with a human IL-4 gene and/or IL-4Rα gene in whole or in part, and methods for making and using the non-human animals, are described. Non-human animals comprising a human or humanized IL-4 gene under control of non-human IL-4 regulatory elements is also provided, including non-human animals that have a replacement of non-human IL-4-encoding sequence with human IL-4-encoding sequence at an endogenous non-human IL-4 locus. Non-human animals comprising a human or humanized IL-4Rα gene under control of non-human IL-4Rα regulatory elements is also provided, including non-human animals that have a replacement of non-human IL-4Rα-encoding sequence with human or humanized IL-4Rα-encoding sequence at an endogenous non-human C IL-4Rα locus. Non-human animals comprising human or humanized IL-4 gene and/or IL-4Rα sequences, wherein the non-human animals are rodents, e.g., mice or rats, are provided. ## Claims ### Claim 1 A triply humanized mouse, comprising (i) a replacement of a genomic DNA of a mouse IL-4Rα gene at an endogenous mouse IL-4Rα locus with a human genomic DNA of a human IL-4Rα gene to form a humanized IL-4Rα gene, wherein the genomic DNA of the mouse IL-4Rα gene comprises the ATG initiation codon of exon 1 through exon 5 of the mouse IL-4Rα gene, and the human genomic DNA of the human IL-4Rα gene comprises the ATG initiation codon of exon 1 through exon 5 of the human IL-4Rα gene, wherein the humanized IL-4Rα gene comprises the ATG initiation codon of exon 1 through exon 5 of the human IL-4Rα gene and exons 6-9 of the mouse IL-4Rα gene, wherein expression of the humanized IL-4Rα gene is under control of the mouse IL-4Rα promoter at the endogenous mouse IL-4Rα locus, (ii) a replacement of a genomic DNA of a mouse IL-4 gene at an endogenous mouse IL-4 locus with a genomic DNA of a human IL-4 gene to form a humanized IL-4 gene, wherein the genomic DNA of the mouse IL-4 gene comprises the ATG initiation codon of exon 1 through exon 4 of the mouse IL-4 gene, and the genomic DNA of the human IL-4 gene comprises the ATG initiation codon of exon 1 through exon 4 of the human IL-4 gene, and wherein expression of the humanized IL-4 gene is under control of the mouse IL-4 promoter at the endogenous mouse IL-4 locus, and (iii) a replacement of a genomic DNA of a mouse IL-33 gene at an endogenous mouse IL-33 locus with a genomic DNA of a human IL-33 gene such that the locus encodes a human IL-33 but not mouse IL-33. ### Claim 2 (depends on 1) The mouse of claim 1, wherein the genomic DNA of the mouse IL-33 gene at the endogenous mouse IL-33 locus being replaced comprises the ATG initiation codon of exon 2 through the stop codon of exon 8 of the mouse IL-33 gene, and the genomic DNA of the human IL-33 gene comprises the ATG initiation codon of exon 2 through exon 8 of the human IL-33 gene. ### Claim 3 (depends on 1) The mouse of claim 1, wherein the mouse is incapable of expressing a mouse IL-4Rα protein, a mouse IL-4 protein, and a mouse IL-33 protein. ### Claim 4 (depends on 2) The mouse of claim 2, wherein the mouse is incapable of expressing a mouse IL-4Rα protein, a mouse IL-4 protein, and a mouse IL-33 protein. ### Claim 5 A mouse embryonic stem cell, comprising (i) a replacement of a genomic DNA of a mouse IL-4Rα gene at an endogenous mouse IL-4Rα locus with a human genomic DNA of a human IL-4Rα gene to form a humanized IL-4Rα gene, wherein the genomic DNA of the mouse IL-4Rα gene comprises the ATG initiation codon of exon 1 through exon 5 of the mouse IL-4Rα gene, and the human genomic DNA of the human IL-4Rα gene comprises the ATG initiation codon of exon 1 through exon 5 of the human IL-4Rα gene, wherein the humanized IL-4Rα gene comprises the ATG initiation codon of exon 1 through exon 5 of the human IL-4Rα gene and exons 6-9 of the mouse IL-4Rα gene, wherein expression of the humanized IL-4Rα gene is under control of the mouse IL-4Rα promoter at the endogenous mouse IL-4Rα locus, (ii) a replacement of a genomic DNA of a mouse IL-4 gene at an endogenous mouse IL-4 locus with a genomic DNA of a human IL-4 gene to form a humanized IL-4 gene, wherein the genomic DNA of the mouse IL-4 gene comprises the ATG initiation codon of exon 1 through exon 4 of the mouse IL-4 gene, and the genomic DNA of the human IL-4 gene comprises the ATG initiation codon of exon 1 through exon 4 of the human IL-4 gene, and wherein expression of the humanized IL-4 gene is under control of the mouse IL-4 promoter at the endogenous mouse IL-4 locus, and (iii) a replacement of a genomic DNA of a mouse IL-33 gene at an endogenous mouse IL-33 locus with a genomic DNA of a human IL-33 gene such that the locus encodes a human IL-33 but not mouse 11-33. ### Claim 6 (depends on 5) A mouse embryo comprising the mouse embryonic stem cell of claim 5. ### Claim 7 (depends on 1) A method, comprising: providing a mouse according to claim 1, administering to said mouse, a candidate human IL-4 or IL-4Rα antagonist and a candidate human IL-33 antagonist, and assessing the pharmacodynamics profiles of said candidate human IL-4 or IL-4Rα antagonist and said candidate human IL-33 antagonist. ### Claim 8 (depends on 7) The method of claim 7, wherein the genomic DNA of the mouse IL-33 gene at the endogenous mouse IL-33 locus being replaced comprises the ATG initiation codon of exon 2 through the stop codon of exon 8 of the mouse IL-33 gene, and the genomic DNA of the human IL-33 gene comprises the ATG initiation codon of exon 2 through exon 8 of the human IL-33 gene. ### Claim 9 (depends on 8) The method of claim 8, wherein the mouse is incapable of expressing a mouse IL-4Rα protein, a mouse IL-4 protein, and a mouse IL-33 protein. ### Claim 10 (depends on 9) The method of claim 9, wherein the candidate human IL-4 or IL-4Rα antagonist is an antibody specific for human IL-4 or IL-4Rα, and the candidate human IL-33 antagonist is an antibody specific for human IL-33. ### Claim 11 (depends on 1) A method, comprising providing a mouse according to claim 1, inducing an inflammatory condition in said mouse, administering to said mouse, a candidate human IL-4 or IL-4Rα antagonist and a candidate human IL-33 antagonist, and determining whether is the inflammatory condition is reduced by the administration. ### Claim 12 (depends on 11) The method of claim 11, wherein the genomic DNA of the mouse IL-33 gene at the endogenous mouse IL-33 locus being replaced comprises the ATG initiation codon of exon 2 through the stop codon of exon 8 of the mouse IL-33 gene, and the genomic DNA of the human IL-33 gene comprises the ATG initiation codon of exon 2 through exon 8 of the human IL-33 gene. ### Claim 13 (depends on 12) The method of claim 12, wherein the mouse is incapable of expressing a mouse IL-4Ra protein, a mouse IL-4 protein, and a mouse IL-33 protein. ### Claim 14 (depends on 13) The method of claim 13, wherein the candidate human IL-4 or IL-4Rα antagonist is an antibody specific for human IL-4 or IL-4Ra, and the candidate human IL-33 antagonist is an antibody specific for human IL-33. ### Claim 15 (depends on 11) The method of claim 11, wherein the inflammatory condition induced is pulmonary inflammation. ### Claim 16 (depends on 15) The method of claim 15, wherein the extent of pulmonary inflammation is determined by measuring (a) mucus accumulation in the airway, (b) eosinophilic infiltrating cells in bronchoalveolar lavage fluid, (c) total circulating IgE, or (d) any combination thereof. ### Claim 17 (depends on 11) The method of claim 11, wherein the inflammatory condition induced is skin inflammation.