Humanized IL-6 and IL-6 receptor

Patent metadata

Field	Value
Publication number	US-9622460-B2
Application number	US-201615177582-A
Country	US
Kind code	B2
Filing date	Jun 9, 2016
Priority date	Oct 28, 2011
Publication date	Apr 18, 2017
Grant date	Apr 18, 2017

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.

First claim

We claim:

1. A genetically modified rodent animal, wherein the genome of said rodent animal comprises a replacement at an endogenous rodent IL-6 locus of a rodent gene encoding IL-6 with a human gene encoding human IL-6, wherein the human gene encoding human IL-6 is under control of endogenous rodent regulatory elements at the endogenous rodent IL-6 locus and is expressed, and the replaced rodent IL-6 gene is not expressed; wherein the genome of said rodent animal further comprises a replacement at an endogenous rodent IL-6Rα locus of a rodent nucleic acid encoding a rodent IL-6Rα ectodomain with a human nucleic acid encoding a human IL-6Rα ectodomain to form a humanized IL-6Rα gene, wherein the humanized IL-6Rα gene is under control of endogenous rodent regulatory elements at the endogenous rodent IL-6Rα locus, wherein the humanized IL-6Rα gene is expressed and encodes a humanized IL-6Rα protein which comprises said human IL-6Rα ectodomain and rodent IL-6Rα transmembrane and cytoplasmic domains; and wherein the rodent animal exhibits an acute phase response to an acute phase inducer and is a rat or a mouse.

2. The genetically modified rodent animal of claim 1 , wherein the rodent animal is a rat.

3. The genetically modified rodent animal of claim 1 , wherein said rodent animal is heterozygous with respect to the replacement at the endogenous rodent IL-6 locus.

4. The genetically modified rodent animal of claim 1 , wherein said rodent animal is homozygous with respect to the replacement at the endogenous rodent IL-6 locus.

5. The genetically modified rodent animal of claim 1 , wherein said rodent animal is heterozygous with respect to said humanized IL-6Rα gene.

6. The genetically modified rodent animal of claim 1 , wherein said rodent animal is homozygous with respect to said humanized IL-6Rα gene.

7. The genetically modified rodent animal of claim 1 , wherein said rodent animal is homozygous with respect to the replacement at the endogenous rodent IL-6 locus, and wherein said rodent animal is homozygous with respect to said humanized IL-6Rα gene.

8. The genetically modified rodent animal of claim 1 , wherein said human gene encoding human IL-6 comprises exon 1 through exon 5 of a human IL-6 gene.

9. A method for making a genetically modified, humanized rodent animal, comprising:
i) replacing an endogenous rodent gene sequence encoding rodent IL-6 at an endogenous rodent IL-6 locus with a human gene encoding human IL-6, and
ii) replacing a nucleic acid encoding the rodent IL-6Rα ectodomain at an endogenous rodent IL-6Rα locus with a human nucleic acid encoding a human IL-6Rα ectodomain to form a humanized IL-6Rα gene, to make a genetically modified, humanized rodent animal whose genome comprises a replacement of the endogenous rodent gene encoding IL-6 and replacement of the nucleic acid encoding the endogenous rodent IL-6 Rα ectodomain with corresponding human sequences,
wherein the human gene encoding human IL-6 is operably linked to endogenous rodent IL-6 regulatory sequences such that the human gene encoding human IL-6 is expressed and the replaced rodent IL-6 gene is not expressed, wherein the humanized IL-6Rα gene is operably linked to endogenous rodent IL-6Rα regulatory sequences such that the humanized IL-6Rα gene is expressed and encodes a humanized IL-6Rα protein that comprises said human IL-6Rα ectodomain and rodent IL-6Rα transmembrane domain and cytoplasmic domain, and wherein the rodent animal exhibits an acute phase response to an acute phase inducer and is a rat or a mouse.

10. The method of claim 9 , wherein said rodent animal is a rat.

11. The method of claim 9 , wherein said humanized rodent animal is heterozygous with respect to the replacement at the endogenous rodent IL-6 locus.

12. The method of claim 9 , wherein said humanized rodent animal is homozygous with respect to the replacement at the endogenous rodent IL-6 locus.

13. The method of claim 9 , wherein said humanized rodent animal is heterozygous with respect to said humanized IL-6Rα gene.

14. The method of claim 9 , wherein said humanized rodent animal is homozygous with respect to said humanized IL-6Rα gene.

15. The method of claim 9 , wherein said humanized rodent animal is homozygous with respect to the replacement at the endogenous rodent IL-6 locus, and wherein said rodent is homozygous with respect to said humanized IL-6Rα gene.

16. The method of claim 9 , wherein said human gene encoding human IL-6 comprises exon 1 through exon 5 of a human IL-6 gene.

Assignees

• Regeneron Pharma

Inventors

• Wang Li-Hsien
• Dore Jr Anthony T
• Stevens Sean
• Murphy Andrew J

Classifications

• C07K14/7155

  for interleukins [IL] · CPC title

• A01K2227/10

  Mammal · CPC title

• C12N15/907

  in mammalian cells · CPC title

• A01K2217/072

  maintaining or altering function, i.e. knock in · CPC title

• A01K2267/02

  Animal zootechnically ameliorated · CPC title