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US 2022/0053742 A1Published application
Humanized il-6 and il-6 receptor
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsLi-Hsien Wang, Anthony T. Dore, Jr., Sean Stevens, Andrew J. Murphy
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Abstract
Mice that comprise a replacement of endogenous mouse IL-6 and/or IL-6 receptor genes are described, and methods for making and using the mice. Mice comprising a replacement at an endogenous IL-6Rα locus of mouse ectodomain-encoding sequence with human ectodomain-encoding sequence is provided. Mice comprising a human IL-6 gene under control of mouse IL-6 regulatory elements is also provided, including mice that have a replacement of mouse IL-6-encoding sequence with human IL-6-encoding sequence at an endogenous mouse IL-6 locus.
Claims17
-20. (canceled)
A method for determining whether a substance induces a human IL-6 mediated acute phase response, comprising
- (a) exposing a genetically modified murine animal to the substance, wherein the genetically modified rodent animal is a mouse or a rat and comprises:
- (i) a humanized IL-6Rα gene at an endogenous murine IL-6Rα locus, wherein the humanized IL-6Rα gene is under control of endogenous murine regulatory sequences at the endogenous murine IL-6Rα locus, and wherein the humanized IL-6Rα gene encodes a humanized IL-6Rα protein that comprises a human IL-6Rα ectodomain, a murine IL-6Rα transmembrane domain and a murine IL-6Rα cytoplasmic domain;
- and
- (ii) a humanized IL-6 gene at an endogenous murine IL-6 locus, wherein the humanized IL-6 gene encodes a human IL-6 protein and is under control of endogenous murine regulatory sequences at the endogenous murine IL-6 locus;
- and
- (b) measuring the level of one or more acute phase response proteins or RNAs, thereby determining that the substance induces a human IL-6 mediated acute phase response based on an increased level of one or more acute phase response proteins or RNAs as a result of the exposure to the substance.
3 more elements
A method for determining whether a substance induces a human IL-6 mediated acute phase response, comprising
- (a) exposing a genetically modified murine animal to the substance in the presence and in the absence of an antagonist of human IL-6Rα, respectively, wherein the genetically modified rodent animal is a mouse or a rat and comprises:
- (i) a humanized IL-6Rα gene at an endogenous murine IL-6Rα locus, wherein the humanized IL-6Rα gene is under control of endogenous murine regulatory sequences at the endogenous murine IL-6Rα locus, and wherein the humanized IL-6Rα gene encodes a humanized IL-6Rα protein that comprises a human IL-6Rα ectodomain, a murine IL-6Rα transmembrane domain and a murine IL-6Rα cytoplasmic domain;
- and
- (ii) a humanized IL-6 gene at an endogenous murine IL-6 locus, wherein the humanized IL-6 gene encodes a human IL-6 protein and is under control of endogenous murine regulatory sequences at the endogenous murine IL-6 locus;
- (b) measuring the level of one or more acute phase response proteins or RNAs;
- and
- (c) determining that the substance induces a human IL-6 mediated acute phase response based on a reduction in a level of an acute phase response protein or RNA in the presence of the human IL-6Rα antagonist as compared to in the absence of the human IL-6Rα antagonist.
4 more elements
A method for determining whether a compound is an antagonist of human IL-6Rα, comprising:
- (a) exposing a genetically modified rodent animal to an acute phase response inducer in the presence and in the absence of the compound, respectively, wherein the genetically modified rodent animal is a mouse or a rat and comprises:
- (i) a humanized IL-6Rα gene at an endogenous murine IL-6Rα locus, wherein the humanized IL-6Rα gene is under control of endogenous murine regulatory sequences at the endogenous murine IL-6Rα locus, and wherein the humanized IL-6Rα gene encodes a humanized IL-6Rα protein that comprises a human IL-6Rα ectodomain, a murine IL-6Rα transmembrane domain and a murine IL-6Rα cytoplasmic domain;
- and
- (ii) a humanized IL-6 gene at an endogenous murine IL-6 locus, wherein the humanized IL-6 gene encodes a human IL-6 protein and is under control of endogenous murine regulatory sequences at the endogenous murine IL-6 locus;
- (b) measuring the level of one or more acute phase response proteins or RNAs;
- and
- (c) determining that compound is a human IL-6Rα antagonist based on a reduction in a level of an acute phase response protein or RNA in the presence of the compound as compared to the absence of the compound.
4 more elements
A targeting nucleic acid construct, comprising
- a human IL-6 nucleotide sequence comprising from ATG in exon 1 through exon 5 and 16 nucleotides downstream of exon 5,
- flanked by mouse IL-6 upstream and downstream homology arms,
- wherein the mouse IL-6 upstream and downstream homology arms mediate integration of the human IL-6 nucleotide sequence into a mouse IL-6 gene locus and replacement of a mouse IL-6 gene sequence, thereby forming a humanized IL-6 gene under control of mouse regulatory sequences at the mouse IL-6 locus.
A targeting nucleic acid construct, comprising
- a human IL-6Rα nucleotide sequence comprising ATG in exon 1 to exon 8 and 69 nucleotides downstream of exon 8,
- flanked by mouse IL-6Rα upstream and downstream homology arms,
- wherein the mouse IL-6Rα upstream and downstream homology arms mediate integration of the human IL-6Rα nucleotide sequence into a mouse IL-6Rα gene locus and replacement of exons 1-8 of the mouse IL-6Rα gene, thereby forming a humanized IL-6Rα gene which is under control of mouse regulatory sequences at the mouse IL-6Rα locus, and which comprises human IL-6Rα exons 1-8 and mouse IL-6Rα exons 9-10.
Dependencies are inferred from the claim text.