US 12,127,537 B2
Genetically modified non-human animals and methods of use thereof
Richard Flavell, Guilford, CT (US); Markus Manz, Zollikon (CH); Anthony Rongvaux, New Haven, CT (US); Till Strowig, Braunschweig (DE); Tim Willinger, Ultran (SE); Andrew J. Murphy, Croton-on-Hudson, NY (US); Sean Stevens, Del Mar, CA (US); and George Yancopoulos, Yorktown Heights, NY (US)
Assigned to Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed by Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US); Yale University, New Haven, CT (US); and Institute for Research in Biomedicine (IRB), Bellinzona (CH)
Filed on May 10, 2021, as Appl. No. 17/316,266.
Application 17/316,266 is a continuation of application No. 16/586,573, filed on Sep. 27, 2019, granted, now 11,026,408.
Application 16/586,573 is a continuation of application No. 15/980,602, filed on May 15, 2018, granted, now 10,433,527, issued on Oct. 8, 2019.
Application 15/980,602 is a continuation of application No. 15/598,080, filed on May 17, 2017, abandoned.
Application 15/598,080 is a continuation of application No. 14/420,318, granted, now 9,820,476, issued on Nov. 21, 2017, previously published as PCT/US2013/058448, filed on Sep. 6, 2013.
Claims priority of provisional application 61/698,002, filed on Sep. 7, 2012.
Claims priority of provisional application 61/775,171, filed on Mar. 8, 2013.
Prior Publication US 2021/0368752 A1, Dec. 2, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. A01K 67/0278 (2024.01); A01K 67/0271 (2024.01); C12N 15/87 (2006.01)
CPC A01K 67/0278 (2013.01) [A01K 67/0271 (2013.01); C12N 15/87 (2013.01); A01K 2207/12 (2013.01); A01K 2207/15 (2013.01); A01K 2217/072 (2013.01); A01K 2217/15 (2013.01); A01K 2227/105 (2013.01); A01K 2267/0331 (2013.01); A01K 2267/0381 (2013.01); A01K 2267/0387 (2013.01)] 18 Claims
 
1. A method for generating a genetically modified, immunodeficient mouse comprising in its genome a replacement of a mouse M-CSF gene with a nucleic acid encoding a human M-CSF polypeptide at a mouse M-CSF gene locus, a replacement of a mouse IL-3 gene with a nucleic acid encoding a human IL-3 polypeptide at a mouse IL-3 gene locus, a replacement of a mouse GM-CSF gene with a nucleic acid encoding a human GM-CSF polypeptide at a mouse GM-CSF gene locus, an insertion of a nucleic acid encoding a human SIRPA polypeptide, and a replacement of a mouse TPO gene with a nucleic acid encoding a human TPO polypeptide at a mouse TPO gene locus, wherein each of the nucleic acids encoding the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the human SIRPA polypeptide, and the human TPO polypeptide is operably linked to a promoter, and wherein the mouse expresses the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the human SIRPA polypeptide, and the human TPO polypeptide, the method comprising:
breeding a first mouse with a second mouse,
wherein the first mouse comprises in its genome a replacement of a mouse M-CSF gene with a nucleic acid encoding a human M-CSF polypeptide at a mouse M-CSF gene locus, a replacement of a mouse IL-3 gene with a nucleic acid encoding a human IL-3 polypeptide at a mouse IL-3 gene locus, a replacement of a mouse GM-CSF gene with a nucleic acid encoding a human GM-CSF polypeptide at a mouse GM-CSF gene locus, and a replacement of a mouse TPO gene with a nucleic acid encoding a human TPO polypeptide at a mouse TPO gene locus, and
the second mouse comprises in its genome an insertion of a nucleic acid encoding a human SIRPA polypeptide, a Rag-2 gene knock-out, and an IL2rg gene knock-out,
wherein each of the nucleic acids encoding the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the human SIRPA polypeptide, and the human TPO polypeptide is operably linked to a promoter, to generate the genetically modified, immunodeficient mouse.