Rodents having a humanized TMPRSS gene
Genetically modified rodents such as mice and rats, and methods and compositions for making and using the same, are provided. The rodents comprise a humanization of at least one endogenous rodent Tmprss gene, such as an endogenous rodent Tmprss2, Tmprss4, or Tmprss11d gene.
1. A rodent whose genome comprises a replacement of a nucleotide sequence of an endogenous rodent Tmprss gene at an endogenous rodent Tmprss locus with a nucleotide sequence of a cognate human TMPRSS gene to form a humanized Tmprss gene,
wherein the humanized Tmprss gene is under control of the promoter of the endogenous rodent Tmprss gene at the endogenous rodent Tmprss locus, and
wherein the humanized Tmprss gene encodes a humanized Tmprss protein that comprises (i) an ectodomain substantially identical to the ectodomain of the human TMPRSS protein encoded by the cognate human TMPRSS gene, and (ii) a cytoplasmic and transmembrane portion that is substantially identical to the cytoplasmic and transmembrane portion of the endogenous rodent Tmprss protein encoded by the endogenous rodent Tmprss gene.
2. The rodent of claim 1 , wherein the rodent is a mouse or a rat.
3. The rodent of claim 1 , wherein the rodent is heterozygous for the humanized Tmprss gene.
4. The rodent of claim 1 , wherein the rodent is homozygous for the humanized Tmprss gene.
5. An isolated rodent cell or tissue whose genome comprises a replacement of a nucleotide sequence of an endogenous rodent Tmprss gene at an endogenous rodent Tmprss locus with a nucleotide sequence of a cognate human TMPRSS gene to form a humanized Tmprss gene,
wherein the humanized Tmprss gene is under control of the promoter of the endogenous rodent Tmprss gene at the endogenous rodent Tmprss locus, and
wherein the humanized Tmprss gene encodes a humanized Tmprss protein that comprises (i) an ectodomain substantially identical to the ectodomain of the human TMPRSS protein encoded by the cognate human TMPRSS gene, and (ii) a cytoplasmic and transmembrane portion that is substantially identical to the cytoplasmic and transmembrane portion of the endogenous rodent Tmprss protein encoded by the endogenous rodent Tmprss gene.
6. The isolated rodent cell or tissue of claim 5 , wherein the rodent cell is a rodent embryonic stem cell.
7. A rodent embryo generated from the rodent embryonic stem cell of claim 6 .
8. A vector comprising a human genomic DNA encoding the ectodomain of a human TMPRSS protein, flanked by a 5′ nucleotide sequence and a 3′ nucleotide sequence that are homologous to genomic DNA sequences flanking a rodent genomic DNA at a rodent Tmprss locus encoding the ectodomain of a cognate rodent Tmprss protein, wherein a homologous recombination-mediated integration of the human genomic DNA into the rodent Tmprss locus results in a replacement of the rodent genomic DNA with the human genomic DNA to form a humanized Tmprss gene which encodes a humanized Tmprss protein that comprises (i) an ectodomain substantially identical to the ectodomain of the human TMPRSS protein encoded by the cognate human TMPRSS gene, and (ii) a cytoplasmic and transmembrane portion that is substantially identical to the cytoplasmic and transmembrane portion of the endogenous rodent Tmprss protein encoded by the endogenous rodent Tmprss gene.
9. A method of providing a rodent whose genome comprises a humanized Tmprss gene, the method comprising:
modifying the genome of a rodent to replace a genomic sequence of an endogenous Tmprss gene at an endogenous rodent Tmprss locus with a genomic sequence of a cognate human TMPRSS gene to form a humanized Tmprss gene,
wherein the humanized Tmprss gene is under control of the promoter of the endogenous rodent Tmprss gene at the endogenous rodent Tmprss locus, and
wherein the humanized Tmprss gene encodes a humanized Tmprss protein that comprises (i) an ectodomain substantially identical to the ectodomain of the human TMPRSS protein encoded by the cognate human TMPRSS gene, and (ii) a cytoplasmic and transmembrane portion that is substantially identical to the cytoplasmic and transmembrane portion of the endogenous rodent Tmprss protein encoded by the endogenous rodent Tmprss gene.
10. A method of making a rodent having a humanized Tmprss gene, comprising:
(a) inserting a genomic fragment into an endogenous rodent Tmprss locus in a rodent embryonic stem cell, said genomic fragment comprising a nucleotide sequence of a cognate human TMPRSS gene, thereby forming a humanized Tmprss gene, wherein the humanized Tmprss gene is under control of the promoter of the rodent Tmprss gene at the endogenous rodent Tmprss locus, and wherein the humanized Tmprss gene encodes a humanized Tmprss protein that comprises (i) an ectodomain substantially identical to the ectodomain of the human TMPRSS protein encoded by the cognate human TMPRSS gene, and (ii) a cytoplasmic and transmembrane portion that is substantially identical to the cytoplasmic and transmembrane portion of the endogenous rodent Tmprss protein encoded by the endogenous rodent Tmprss gene;
(b) obtaining a rodent embryonic stem cell comprising the humanized Tmprss gene of (a); and,
(c) creating a rodent using the rodent embryonic stem cell of (b).
11. A method of assessing the therapeutic efficacy of a compound in treating influenza virus infection, comprising:
administering an influenza virus and a candidate compound to the rodent of claim 1 ; and
monitoring the presence and severity of influenza virus infection in the rodent to determine the therapeutic efficacy of the candidate compound.
12. The rodent of claim 1 , wherein the rodent is a mouse.
13. The isolated rodent cell or tissue of claim 6 , wherein the rodent cell is a mouse or rat embryonic stem cell.
14. The isolated rodent cell or tissue of claim 6 , wherein the rodent cell is a mouse embryonic stem cell.
15. The rodent embryo of claim 7 , wherein the rodent embryo is a mouse or rat embryo.
16. The rodent embryo of claim 7 , wherein the rodent embryo is a mouse embryo.
17. The method of claim 9 , wherein the rodent is a mouse or a rat.
18. The method of claim 9 , wherein the rodent is a mouse.
19. The method of claim 10 , wherein the rodent is a mouse or a rat.
20. The method of claim 10 , wherein the rodent is a mouse.
21. The method of claim 11 , wherein the rodent is a mouse or a rat.
22. The method of claim 11 , wherein the rodent is a mouse.
23. The method of claim 11 , wherein the candidate compound is an antibody or antigen-binding fragment thereof specific for a human TMPRSS protein.