Genetically modified major histocompatibility complex mice

a histocompatibility complex and mouse technology, applied in the field of genetically modified nonhuman animals, can solve problems such as the destruction of cells presenting such peptides

Inactive Publication Date: 2015-12-03
REGENERON PHARM INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0011]A biological system for generating or identifying peptides that associate with human MHC class I proteins and chimeras thereof and bind CD8+T cells, as well as peptides that associate with human MHC class II proteins and chimeras thereof and bind to CD4+T cells, is provided. Non-human animals comprising non-human cells that express humanized molecules that function in the cellular immune response are provided. Humanized rodent loci that encode humanized MHC I and MHC II proteins are also provided. Humanized rodent cells that express humanized MHC molecules are also provided. In vivo and in vitro systems are provided that comprise humanized rodent cells, wherein the rodent cells express one or more humanized immune system molecules.

Problems solved by technology

However, in some diseases (e.g., cancer, autoimmune diseases) peptides derived from self-proteins become the target of the cellular component of the immune system, which results in destruction of cells presenting such peptides.

Method used

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  • Genetically modified major histocompatibility complex mice
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  • Genetically modified major histocompatibility complex mice

Examples

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example 1

Engineering a Mouse Comprising Humanized MHC I and MHC II Loci

[0100]The various steps involved in engineering a mouse comprising humanized MHC I and MHC II loci, with corresponding and additional endogenous MHC I and MHC II loci deletions (HLA-A2 / H-2K, HLA-DR2 / H-2E, H-2A-del, H-2D-del) are depicted in FIG. 4. Detailed description of the steps appears below.

example 1.1

Engineering Mouse ES Cells Comprising Humanized MHC I Gene

[0101]The mouse H-2K gene was humanized in a single step by construction of a unique targeting vector from human and mouse bacterial artificial chromosome (BAC) DNA using VELOCIGENE® technology (see, e.g., U.S. Pat. No. 6,586,251 and Valenzuela et al. (2003) High-throughput engineering of the mouse genome coupled with high-resolution expression analysis. Nat. Biotech. 21(6): 652-659). DNA from mouse BAC clone RP23-173k21 (Invitrogen) was modified by homologous recombination to replace the genomic DNA encoding the α1, α2 and α3 domains of the mouse H-2K gene with human genomic DNA encoding the α1, α2 and α3 subunits of the human HLA-A2 gene (FIG. 5). Detailed steps for construction of BAC DNA can be found in U.S. Patent Application Publication No. 2013-0111617, incorporated herein by reference. The targeted BAC DNA was used to electroporate mouse F1H4 ES cells to create modified ES cells for generating mice that express a chim...

example 1.2

Engineering Mouse ES Cells Comprising Deletion of MHC II Loci

[0103]The targeting vector for introducing a deletion of the endogenous MHC class II H-2Ab1, H-2Aa, H-2Eb1, H-2Eb2, and H-2Ea genes was made using VELOCIGENE® genetic engineering technology (see, e.g., U.S. Pat. No. 6,586,251 and Valenzuela et al., supra). Bacterial Artificial Chromosome (BAC) RP23-458i22 (Invitrogen) DNA was modified to delete the endogenous MHC class II genes H-2Ab1, H-2Aa, H-2Eb1, H-2Eb2, and H-2Ea.

[0104]Briefly, upstream and downstream homology arms were derived by PCR of mouse BAC DNA from locations 5′ of the H-2Ab1 gene and 3′ of the H-2Ea gene, respectively. These homology arms were used to make a cassette that deleted ˜79 kb of RP23-458i22 comprising genes H-2Ab1, H-2Aa, H-2Eb1, H-2Eb2, and H-2Ea of the MHC class II locus by bacterial homologous recombination (BHR). This region was replaced with a neomycin cassette flanked by lox2372 sites. The final targeting vector included from 5′ to 3′ a 26 kb ...

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Abstract

The invention provides genetically modified non-human animals that express chimeric human/non-human MHC I and MHC II polypeptides and/or human or humanized β2 microglobulin polypeptide, as well as embryos, cells, and tissues comprising the same. Also provided are constructs for making said genetically modified animals and methods of making the same. Methods of using the genetically modified animals to study various aspects of human immune system are provided.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application claims benefit under 35 U.S.C. §119(e) of U.S. provisional patent application Ser. No. 62 / 039,333, filed Aug. 19, 2014 and is a continuation-in-part application of U.S. application Ser. No. 14 / 185,316, filed Feb. 20, 2014, which claims the benefit under 35 §119(e) of U.S. Provisional Patent Application Ser. No. 61 / 767,811, filed Feb. 22, 2013, all of which are hereby incorporated by reference in their entirety.FIELD OF THE INVENTION[0002]The present invention relates to a genetically modified non-human animal, e.g., a rodent (e.g., a mouse or a rat), that expresses a human or humanized Major Histocompatibility Complex (MHC) class I and a human or humanized MHC class II molecules. The invention also relates to a genetically modified non-human animal, e.g., a mouse or a rat, that expresses a human or humanized MHC I protein (e.g., MHC I α chain) and a human or humanized MHC II protein (e.g., MHC II α and MHC II β chains), a...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01K67/027
CPCA01K67/0271A01K2217/072A01K2227/105A01K2207/15A01K67/0278C07K14/70514C07K14/70517C07K14/70539C12N15/8509A01K2267/03
Inventor VORONINA, VERAGURER, CAGANMURPHY, ANDREW J.MACDONALD, LYNNXUE, YINGZE
Owner REGENERON PHARM INC
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