US 12,433,265 B2
Non-human animals having a humanized a proliferation-inducing ligand gene
John McWhirter, Hastings-on-Hudson, NY (US); Cagan Gurer, Chappaqua, NY (US); Lynn Macdonald, Harrison, NY (US); and Andrew J. Murphy, Croton-on-Hudson, NY (US)
Assigned to Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed by Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed on Feb. 5, 2021, as Appl. No. 17/168,256.
Application 17/168,256 is a division of application No. 16/001,070, filed on Jun. 6, 2018, granted, now 10,952,416.
Application 16/001,070 is a division of application No. 14/537,320, filed on Nov. 10, 2014, abandoned.
Claims priority of provisional application 61/905,986, filed on Nov. 19, 2013.
Prior Publication US 2021/0161113 A1, Jun. 3, 2021
Int. Cl. A01K 67/0278 (2024.01); C07K 14/47 (2006.01); C07K 14/705 (2006.01); C12N 15/85 (2006.01); G01N 33/50 (2006.01); A01K 67/0271 (2024.01)
CPC A01K 67/0278 (2013.01) [C07K 14/47 (2013.01); C07K 14/70575 (2013.01); C12N 15/8509 (2013.01); G01N 33/5005 (2013.01); A01K 67/0271 (2013.01); A01K 2207/12 (2013.01); A01K 2207/15 (2013.01); A01K 2217/052 (2013.01); A01K 2217/07 (2013.01); A01K 2217/072 (2013.01); A01K 2227/105 (2013.01); A01K 2267/03 (2013.01); A01K 2267/0387 (2013.01); C07K 2319/00 (2013.01); C12N 2015/8527 (2013.01)] 8 Claims
 
1. A method of engrafting human cells into a genetically modified mouse, the method comprising steps of:
(a) providing a mouse whose genome comprises
(i) a recombination activating gene 2 (Rag2) gene knock-out and an IL2 receptor gamma chain (IL2rg) gene knock-out, and
(ii) a replacement of a mouse genomic fragment comprising exons 2-5 of an endogenous mouse April gene at an endogenous mouse April locus with a human genomic fragment comprising exons 2-6 of a human APRIL gene to form a humanized April gene,
wherein the humanized April gene is under control of a mouse April promoter at the endogenous mouse April locus and encodes a humanized April protein comprising an extracellular portion of the human APRIL protein encoded by the human APRIL gene linked to an intracellular portion of the mouse April protein encoded by the endogenous mouse April gene;
wherein the coding exons of the humanized April gene consist of exon 1 of the endogenous mouse April gene and exons 2-6 of the human APRIL gene;
wherein the humanized April gene comprises a 5′ portion of intron 1 of the endogenous mouse April gene, wherein said 5′ portion consists of 100 bp of intron 1 of the endogenous mouse April gene; and
wherein the genetically modified mouse expresses the humanized April protein; and
(b) transplanting the human cells into the genetically modified mouse, wherein the human cells are human hematopoietic stem cells or human B cells.