Genetically modified non-human animal with human or chimeric PD-L1

Patent metadata

Field	Value
Publication number	US-10945418-B2
Application number	US-201916428941-A
Country	US
Kind code	B2
Filing date	May 31, 2019
Priority date	Aug 31, 2016
Publication date	Mar 16, 2021
Grant date	Mar 16, 2021

Abstract

This disclosure relates to genetically modified animal expressing human or chimeric (e.g., humanized) programmed death-ligand 1 (PD-L1, PDL1, or B7-H1), and methods of use thereof. In one aspect, the disclosure relates to genetically-modified, non-human animals whose genome comprises at least one chromosome comprising a sequence encoding a human or chimeric programmed death-ligand 1 (PD-L1).

First claim

What is claimed is:

1. A genetically-modified mouse whose genome comprises at least one chromosome comprising a nucleotide sequence encoding a chimeric programmed death-ligand 1 (PD-L1) at an endogenous PD-L1 gene locus, wherein the chimeric PD-L1 comprises SEQ ID NO: 33, wherein the mouse expresses the chimeric PD-L1 on the surface of one or more spleen cells after the genetically modified mouse is challenged by anti-CD3 antibody.

2. The mouse of claim 1 , wherein the sequence encoding the chimeric PD-L1 is operably linked to an endogenous regulatory element at the endogenous PD-L1 gene locus in the at least one chromosome.

3. The mouse of claim 1 , wherein the mouse does not express endogenous PD-L1.

4. The mouse of claim 1 , wherein the mouse is homozygous with respect to the sequence encoding the chimeric PD-L1 at the endogenous PD-L1 gene locus.

5. The mouse of claim 1 , wherein the mouse whose genome further comprises a nucleotide sequence encoding a humanized PD-1 protein at the endogenous PD-1 gene locus, wherein the mouse expresses the humanized PD-1 protein on the surface of one or more spleen cell after the genetically modified mouse is challenged by anti-CD3 antibody.

6. The mouse of claim 1 , wherein exon 3 at the endogenous PD-L1 gene locus is modified by CRISPR with sgRNAs that target SEQ ID NO: 1 and SEQ ID NO: 11.

7. A genetically modified mouse or a progeny thereof, wherein the genetically modified mouse is made by a method comprising the steps of: modifying genome of an embryo of a mouse by CRISPR with sgRNAs that target SEQ ID NO: 1 and SEQ ID NO: 11 to introduce a fragment of human PD-L1 gene, wherein an endogenous PD-L1 gene locus in the genome of the embryo is modified; and transplanting the embryo to a recipient mouse to produce the genetically-modified mouse, wherein the genetically-modified mouse expresses a chimeric PD-L1 comprising a fragment of human PD-L1 protein on the surface of one or more spleen cells after the genetically-modified mouse is challenged by anti-CD3 antibody.

8. The genetically-modified mouse or the progeny thereof of claim 7 , wherein the mouse has a C57BL/6 background.

9. A genetically-modified mouse whose genome comprises at least one chromosome comprising a sequence encoding a human PD-L1 at an endogenous PD-L1 gene locus, wherein the sequence encoding the human PD-L1 is operably linked to an endogenous regulatory element at the endogenous PD-L1 gene locus in the at least one chromosome, wherein the mouse expresses a human PD-L1 on the surface of one or more spleen cells after the genetically modified mouse is challenged by anti-CD3 antibody.

10. The mouse of claim 9 , wherein the mouse does not express endogenous PD-L1.

11. The mouse of claim 9 , wherein the mouse is homozygous with respect to the sequence encoding the human PD-L1 at the endogenous PD-L1 gene locus.

Assignees

• Biocytogen Pharmaceuticals Beijing Co Ltd

Inventors

• Shen Yuelei
• Bai Yang
• Guo Yanan
• Huang Rui
• Zhou Xiaofei
• Guo Chaoshe

Classifications

• C12N15/8509 Primary

  for producing genetically modified animals, e.g. transgenic · CPC title

• C07H21/04

  with deoxyribosyl as saccharide radical · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• A01K2267/0331

  Animal model for proliferative diseases · CPC title

• A01K2207/15

  Humanized animals · CPC title