US 11,716,974 B2
Human liver chimeric mouse with deficient P450 oxidoreductase
Karl-Dimiter Bissig, Houston, TX (US); Maria de las Mercedes Barzi Dieguez, Houston, TX (US); and Peter Francis Pankowicz, Houston, TX (US)
Assigned to Baylor College of Medicine, Houston, TX (US)
Appl. No. 16/309,871
Filed by Baylor College of Medicine, Houston, TX (US)
PCT Filed Jun. 27, 2017, PCT No. PCT/US2017/039474
§ 371(c)(1), (2) Date Dec. 13, 2018,
PCT Pub. No. WO2018/005471, PCT Pub. Date Jan. 4, 2018.
Claims priority of provisional application 62/509,942, filed on May 23, 2017.
Claims priority of provisional application 62/355,102, filed on Jun. 27, 2016.
Prior Publication US 2019/0166810 A1, Jun. 6, 2019
Int. Cl. A01K 67/027 (2006.01); A61K 49/00 (2006.01)
CPC A01K 67/0271 (2013.01) [A01K 67/0276 (2013.01); A61K 49/0008 (2013.01); A01K 2207/15 (2013.01); A01K 2217/075 (2013.01); A01K 2217/206 (2013.01)] 1 Claim
 
1. A method for preparing a chimeric mouse comprising human hepatocytes, the method comprising:
(a) providing a transgenic mouse whose genome comprises:
(i) a liver specific homozygous deletion of endogenous NADPH-P450 oxidoreductase (Por) gene, wherein the deleted Por gene is a conditional knockout of the endogenous Por gene such that no functional endogenous Por protein is produced; and
(ii) a homozygous deletion of endogenous Rag2, IL-2Rg and Fah genes (Fah−/− Rag2−/−IL-2rg−/−), wherein the mouse is immune-deficient, lacking T, B and NK cells,
wherein the transgenic mouse having a liver specific homozygous deletion of endogenous Por gene of (i) is generated by:
1) providing a mouse comprising a floxed allele of the endogenous Por gene with at least one dose of an adenoviral vector encoding Cre recombinase (Ade-Cre), wherein the at least a first dose is sufficient such that no functional endogenous Por protein is produced; or
2) crossing a first transgenic mouse comprising a foxed allele of the endogenous Por gene with a second transgenic mouse strain that expresses CRE recombinase under the liver-specific promoter,
and wherein the liver of the transgenic mouse accumulated lipid as compared to a wild type mouse; and
(b) transplanting human hepatocytes into the transgenic mouse of step (a) to produce a chimeric Rag−/−Il-2Rg−/− Fah−/− Por−/− mouse, wherein the human hepatocytes account for at least 70% of all hepatocytes in the liver of the chimeric Rag−/−Il-2Rg−/− Fah−/− Por−/− mouse.