Genetically modified mice having a humanized programmed cell death 1 gene
Non-human animals, and methods and compositions for making and using the same, are provided, wherein the non-human animals comprise a humanization of a Programmed cell death 1 (Pdcd1) gene. The non-human animals, in some embodiments, comprise a genetic modification to an endogenous Pdcd1 gene so that the non-human animals express a PD-1 polypeptide that includes a human portion and an endogenous portion (e.g., a non-human portion).
1 . A genetically modified mouse embryonic stem(ES) cell whose genome comprises a humanized Programmed cell death 1 (Pdcd1) gene comprising a replacement of exon 2 and at least part of exon 3 of an endogenous Pdcd1 gene with exon 2 and at least part of exon 3 of a human Pdcd1 gene, wherein expression of the humanized Pdcd1 gene is under the control of an endogenous Pdcd1 promoter, and the humanized Pdcd1 gene encodes a humanized PD-1 polypeptide comprising an N-terminal immunoglobulin variable (V) domain of a human PD-1 polypeptide and an intracellular domain of a mouse PD-1 polypeptide.
2 . The mouse ES cell of claim 1 , wherein the humanized PD-1 polypeptide comprises a mouse signal peptide.
3 . The mouse ES cell of claim 1 , wherein the humanized PD-1 polypeptide comprises the transmembrane sequence of the endogenous mouse PD-1 polypeptide.
4 . The mouse ES cell of claim 1 , wherein the humanized PD-1 polypeptide comprises amino acids 26-169 of a human PD-1 polypeptide.
5 . The genetically modified mouse ES cell of claim 1 , wherein the humanized Pdcd1 gene comprises exon 1, exon 4, exon 5, and at least part of exon 3 of the endogenous Pdcd1 gene.
6 . The mouse ES cell of claim 1 , wherein the mouse ES cell is homozygous for the humanized Pdcd1 gene.
7 . A mouse embryo comprising the mouse ES cell of claim 1 .
8 . The mouse embryo of claim 7 , wherein the humanized PD-1 polypeptide comprises a mouse signal peptide.
9 . The mouse embryo of claim 7 , wherein the humanized PD-1 polypeptide comprises the transmembrane sequence of the endogenous mouse PD-1 polypeptide.
10 . The mouse embryo of claim 7 , wherein the humanized PD-1 polypeptide comprises amino acids 26-169 of a human PD-1 polypeptide.
11 . The mouse embryo of claim 7 , wherein the humanized Pdcd1 gene comprises exon 1, exon 4, exon 5, and at least part of exon 3 of the endogenous Pdcd1 gene.
12 . The mouse embryo of claim 7 , wherein the mouse ES cell is homozygous for the humanized Pdcd1 gene.
13 . A method of making a genetically modified mouse, the method comprising:
a) introducing the genetically modified mouse ES cell of claim 1 into a mouse embryo;
b) implanting the mouse embryo obtained in step a) into a female mouse for gestation; and
c) obtaining a genetically modified mouse whose genome comprises said humanized Pdcd1 gene, wherein the genetically modified mouse expresses said humanized PD-1 polypeptide.
14 . A method of making a genetically modified mouse embryonic stem(ES) cell, the method comprising:
replacing exon 2 and at least part of exon 3 of an endogenous Programmed cell death 1 (Pdcd1) gene in a mouse ES cell with exon 2 and at least part of exon 3 of a human Pdcd1 gene such that a genetically modified mouse ES cell is obtained whose genome comprises a humanized Programmed cell death 1 (Pdcd1) gene comprising a replacement of exon 2 and at least part of exon 3 of the endogenous Pdcd1 gene with exon 2 and at least part of exon 3 of the human Pdcd1 gene, wherein expression of the humanized Pdcd1 gene is under the control of an endogenous Pdcd1 promoter, and the humanized Pdcd1 gene encodes a humanized PD-1 polypeptide comprising an N-terminal immunoglobulin variable (V) domain of a human PD-1 polypeptide and an intracellular domain of a mouse PD-1 polypeptide.
15 . The method of claim 14 , wherein the humanized PD-1 polypeptide comprises a mouse signal peptide.
16 . The method of claim 14 , wherein the humanized PD-1 polypeptide comprises the transmembrane sequence of the endogenous mouse PD-1 polypeptide.
17 . The method of claim 14 , wherein the humanized PD-1 polypeptide comprises amino acids 26-169 of a human PD-1 polypeptide.
18 . The method of claim 14 , wherein the humanized Pdcd1 gene comprises exon 1, exon 4, exon 5, and at least part of exon 3 of the endogenous Pdcd1 gene.
19 . The method of claim 14 , wherein the mouse ES cell is homozygous for the humanized Pdcd1 gene.