Humanized M-CSF mice

Patent metadata

Field	Value
Publication number	US-9655352-B2
Application number	US-201414469308-A
Country	US
Kind code	B2
Filing date	Aug 26, 2014
Priority date	Feb 15, 2011
Publication date	May 23, 2017
Grant date	May 23, 2017

Abstract

Genetically modified mice comprising a nucleic acid sequence encoding a human M-CSF protein are provided. Also provided are genetically modified mice comprising a nucleic acid sequence encoding a human M-CSF protein that have been engrafted with human cells such as human hematopoietic cells, and methods for making such engrafted mice. These mice find use in a number of applications, such as in modeling human immune disease and pathogen infection; in in vivo screens for agents that modulate hematopoietic cell development and/or activity, e.g. in a healthy or a diseased state; in in vivo screens for agents that are toxic to hematopoietic cells; in in vivo screens for agents that prevent against, mitigate, or reverse the toxic effects of toxic agents on hematopoietic cells; in in vivo screens of human hematopoietic cells from an individual to predict the responsiveness of an individual to a disease therapy, etc.

First claim

We claim:

1. A method of making a humanized Macrophage Colony Stimulating Factor (M-CSF) mouse, the method comprising:
transforming a mouse embryonic stem cell with a nucleic acid sequence comprising a coding sequence for a human M-CSF protein,
wherein the coding sequence for the human M-CSF protein integrates into the genome of the mouse embryonic stem cell, replacing the endogenous M-CSF gene, and wherein the integrated coding sequence is operably linked to the endogenous promoter of the mouse M-CSF gene;
introducing the transformed mouse embryonic stem cell into a mouse embryo; and
generating a humanized M-CSF mouse from the mouse embryo, wherein the humanized M-CSF mouse comprises the coding sequence for the human M-CSF protein incorporated into its genome and operably linked to the endogenous promoter of the mouse M-CSF gene

wherein the humanized M-CSF mouse comprises a Recombination Activating Gene 2 (Rag2) gene knock-out and an Interleukin 2 Receptor, Gamma Chain (IL2rg) gene knock-out, and wherein the humanized M-CSF mouse expresses M-CSF RNA encoded by the coding sequence in bone marrow, spleen, blood, liver, brain, lung, testis and kidney.

2. The method of claim 1 , wherein the humanized M-CSF mouse comprises two copies of the coding sequence.

3. The method of claim 1 , wherein the humanized M-CSF mouse comprises a null mutation in at least one mouse M-CSF allele and the null mutation is a deletion of mouse M-CSF exons 2-9.

4. The method of claim 1 , further comprising transplanting human cells into the humanized M-CSF mouse.

5. The method of claim 4 , wherein the human cells are hematopoietic cells.

6. The method of claim 4 , further comprising infecting the humanized M-CSF mouse with a human pathogen.

Assignees

• Regeneron Pharma
• Univ Yale
• Inst For Res In Biomedicine (Irb)

Inventors

• Murphy Andrew J
• Stevens Sean
• Rathinam Chozhavendan
• Eynon Elizabeth
• Manz Markus
• Flavell Richard
• Yancopoulos George D

Classifications

• A01K2217/15

  Animals comprising multiple alterations of the genome, by transgenesis or homologous recombination, e.g. obtained by cross-breeding · CPC title

• A01K2267/0337

  Animal models for infectious diseases · CPC title

• A01K67/0278 Primary

  Knock-in vertebrates, e.g. humanised vertebrates · CPC title

• A01K2227/105

  Murine · CPC title

• C12N15/8509 Primary

  for producing genetically modified animals, e.g. transgenic · CPC title