US 11,882,816 B2
Genetically engineered non-human mammal, construction method therefor and use thereof
Miao Wang, Beijing (CN); and Hong Chen, Beijing (CN)
Assigned to Miao Wang, Beijing (CN)
Appl. No. 16/609,110
Filed by Miao Wang, Beijing (CN)
PCT Filed Apr. 28, 2018, PCT No. PCT/CN2018/085097
§ 371(c)(1), (2) Date Oct. 28, 2019,
PCT Pub. No. WO2018/196874, PCT Pub. Date Nov. 1, 2018.
Claims priority of application No. 201710292460.X (CN), filed on Apr. 28, 2017.
Prior Publication US 2020/0077632 A1, Mar. 12, 2020
Int. Cl. A01K 67/027 (2006.01); C12N 15/90 (2006.01)
CPC A01K 67/0276 (2013.01) [C12N 15/90 (2013.01); C12N 2310/20 (2017.05); C12N 2517/02 (2013.01); C12N 2800/107 (2013.01); C12N 2800/60 (2013.01)] 9 Claims
 
1. A genetically engineered non-human mammal, wherein the non-human mammal has a genome with the ApoE gene therein being disrupted so that ApoE is not expressed, and wherein the genome comprises (i) an exogenous nucleotide sequence encoding an siRNA against scavenger receptor class B type I (SR-BI), the exogenous nucleotide sequence being operably linked to an ApoE gene promoter, and (ii) one or more exogenous nucleotide sequences encoding a vasoconstrictor operably linked to an inducible promoter,
wherein the exogenous nucleotide sequence encoding the siRNA and the one or more exogenous nucleotide sequences encoding the vasoconstrictor are inserted into the ApoE gene, and wherein the inserted exogeneous nucleotide sequence encoding the siRNA and the inserted one or more exogenous nucleotide sequences encoding the vasoconstrictor disrupt the ApoE gene so that ApoE is not expressed,
wherein expression of the siRNA knocks down SR-BI expression in the liver of the non-human mammal,
wherein the coding sequence of the siRNA has a homology of 100% to the sequence as shown in SEQ ID NO. 4,
wherein the vasoconstrictor is angiotensin II,
wherein the inducible promoter is a chemical-inducible promoter, and the chemical inducible promoter is a tetracycline-inducible promoter, and
wherein the homozygous form of the genetically engineered non-human mammal displays hypercholesterolemia, hyperlipidemia, increase arteriosclerotic lesion formation, and hypertension as compared to the genetically engineered non-human mammal comprising a disruption of the ApoE gene so that ApoE is not expressed.