Murine-mhc-deficient hla-transgenic nod-mouse models for t1d therapy development

Abstract

The present disclosure relates to genetically modified non-obese diabetic (NOD) mice deficient in murine class I MHC molecules, class II molecules, or both class I and class II MHC molecules. The MHC knockout transgenic mice provided herein are useful, for example, for developing therapies for diabetes.

1 . A genetically modified non-obese diabetic (NOD) mouse comprising in the genome of the NOD mouse a mutation in a gene encoding H2-K and/or a mutation in a gene encoding H2-D. 
     
     
         2 . The genetically modified NOD mouse of  claim 1  comprising in the genome of the NOD mouse a mutation in a gene encoding H2-K and a mutation in a gene encoding H2-D. 
     
     
         3 . The genetically modified NOD mouse of  claim 1 , wherein the genome of the NOD mouse comprises a homozygous mutation the gene encoding H2-K. 
     
     
         4 . The genetically modified NOD mouse of  claim 1 , wherein the genome of the NOD mouse comprises a homozygous mutation in the gene encoding H2-D. 
     
     
         5 . The genetically modified NOD mouse of  claim 1 , wherein cells of the mouse do not express H2-K and/or do not express H2-D. 
     
     
         6 . The genetically modified NOD mouse of  claim 1 , wherein the gene encoding H2-K is H2-K1 d  and/or the gene encoding H2-D is H2-D1 b . 
     
     
         7 . The genetically modified NOD mouse of  claim 1  further comprising in the genome of the NOD mouse a mutation in a gene encoding H2-A. 
     
     
         8 . The genetically modified NOD mouse of  claim 7 , wherein the genome of the NOD mouse comprises a homozygous mutation in the genes encoding H2-A. 
     
     
         9 . The genetically modified NOD mouse of  claim 7 , wherein cells of the mouse do not express H2-A. 
     
     
         10 . The genetically modified NOD mouse of  claim 7 , wherein the gene encoding H2-A is H2-Ab1 g7 . 
     
     
         11 . The genetically modified NOD mouse of  claim 7 , wherein the genome of the NOD mouse further comprises a nucleic acid encoding human HLA-A and/or a nucleic acid encoding human HLA-B 
     
     
         12 . The genetically modified NOD mouse of  claim 11 , wherein the HLA-A is HLA-A*02:01 (HLA-A2) and/or the HLA-B is HLA-B*39:06 (HLA-B39). 
     
     
         13 . The genetically modified NOD mouse of  claim 1 , wherein the genome of the NOD mouse further comprises a nucleic acid encoding human HLA-DQ and/or a nucleic acid encoding human HLA-DR. 
     
     
         14 . The genetically modified NOD mouse of  claim 14 , wherein the HLA-DQ is HLA-DQ8 and/or the HLA-DR is HLA-DR3/4. 
     
     
         15 . A genetically modified non-obese diabetic (NOD) mouse comprising in the genome of the NOD mouse:
 a homozygous mutation in H2-D1 b , a homozygous mutation in H2-K1 d , and a homozygous mutation in H2-Ab1 g7 , optionally designated NOD/ShiLtDvs-H2-K1 em1Dvs  H2-Ab1 em1Dvs  H2-D1 em5Dvs /Dvs;   a homozygous mutation in H2-D1 b , a homozygous mutation in H2-K1 d , and a human HLA-A2 transgene, optionally designated NOD/ShiLtDvs-H2-K1 em1Dvs  H2-D1 em5Dvs  Tg(HLA-A/H2-D/B2M)1Dvs/Dvs; or   a homozygous mutation in H2-D1 b , a homozygous mutation in H2-K1 d , and a human HLA-B39 transgene, optionally designated NOD/ShiLtDvs-H2-K1 em1Dvs  H2-D1 em5Dvs  Tg(HLA-B39/H2-D/B2m)2Dvs/Dvs.   
     
     
         16 .- 17 . (canceled) 
     
     
         18 . A method of producing the genetically modified NOD mouse of any one of  claim 1  using CRISPR/Cas genome editing to introduce at least one of the mutations into the genome of the NOD mouse. 
     
     
         19 . A cell comprising in the genome of the cell a homozygous mutation in a gene encoding H2-K and/or a homozygous mutation in a gene encoding H2-D. 
     
     
         20 .- 21 . (canceled) 
     
     
         22 . A rodent comprising the cell of  claim 19 . 
     
     
         23 . A method comprising administering a test agent to the genetically modified NOD mouse of  claim 1 , and assaying the genetically modified NOD mouse for a symptom of diabetes. 
     
     
         24 . A method comprising introducing into the genome of the genetically modified NOD mouse of  claim 1  a nucleic acid encoding a human MHC class I molecule and/or a nucleic acid encoding a human MHC class II molecule, and producing a humanized NOD mouse comprising a genome that expresses the human MHC class I molecule and/or expresses the human MHC class II molecule. 
     
     
         25 .- 28 . (canceled)