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Construction method of IL-13 gene humanized non-human animal and application thereof

A technology of non-human animals and construction methods, which is applied in the fields of botany equipment and methods, biochemical equipment and methods, applications, etc., and can solve problems such as screening and evaluating drug efficacy, failure to identify mice, and failure to use ordinary mice

Active Publication Date: 2020-11-13
BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the amino acid sequence of human IL-13 is significantly different from the corresponding protein in rodents, the identity is only 59%, therefore, the antibody that recognizes human IL-13 protein usually cannot recognize mouse IL-13, that is, it cannot be detected by ordinary small Rats to Screen and Evaluate the Efficacy of Drugs Targeting IL-13

Method used

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  • Construction method of IL-13 gene humanized non-human animal and application thereof
  • Construction method of IL-13 gene humanized non-human animal and application thereof
  • Construction method of IL-13 gene humanized non-human animal and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0140] Example 1 Preparation of IL-13 Gene Humanized Mice

[0141] Mouse IL-13 gene (NCBI Gene ID: 16163, located at position 53631323 to 53634702 of chromosome 11 NC_000077.6, based on transcript NM_008355.3 and its encoded protein NP_032381.1 (SEQ ID NO: 1)) and human IL-13 gene (NCBI Gene ID: 3596, located at position 132656522 to 132661110 of chromosome 5 NC_000005.10, based on the transcript NM_002188.3 and its encoded protein NP_002179.2 (SEQ ID NO: 2)) comparison schematic diagram as shown figure 1 shown.

[0142] In order to achieve the purpose of the present invention, the gene sequence encoding human IL-13 protein can be introduced into the mouse endogenous IL-13 gene locus, so that the mouse expresses humanized IL-13 protein. Specifically, the specific mouse IL-13 gene sequence can be replaced with the sequence of the human IL-13 gene at the mouse endogenous IL-13 gene locus by gene editing technology, such as at least containing the mouse IL-13 gene. Partial sequ...

Embodiment 2

[0163] Example 2 Asthma model induced by ovalbumin (OVA) combined with aluminum hydroxide

[0164] IL-13 gene humanized mice (5-8 weeks) were selected and sensitized by intraperitoneal injection of ovalbumin (OVA) combined with aluminum hydroxide for 3 times. Induce and make asthma model (modeling scheme see Figure 8 ), the control group replaced OVA with PBS, and samples were obtained on day 26 for analysis. The G1 group was the blank control group, the G2 and G3 groups were the modeling groups, and the G4, G5 and G6 groups were the administration groups. After the injection sensitization, the anti-human IL-13 antibody Lebrikizumab or tralokinumab was intraperitoneally injected according to different administration regimens ( Dosing regimen Table 2).

[0165] Figure 9 The test results showed that leukocytes (CD45+ cells) in bronchoalveolar lavage fluid (BALF) ( Figure 9 A) and eosinophils (Eos) ( Figure 9 B) The number in the modeling group (G2) was slightly higher t...

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Abstract

The invention provides a construction method of an IL-13 gene humanized non-human animal of the IL-13 gene humanized non-human animal and an application in the field of biological medicine. The invention further provides a targeting vector, a humanized IL-13 gene, an IL-13 gene humanized cell and an application of the IL-13 gene humanized cell. According to the non-human animal obtained through the construction method, the human IL-13 protein specifically combined with the human IL-13 antibody can be normally expressed in vivo, and the non-human animal can be further used for preparing a disease animal model and evaluating and screening the in-vivo drug effect of the human IL-13 antibody.

Description

technical field [0001] The invention belongs to the field of animal genetic engineering and genetic modification, and in particular relates to a non-human animal with humanized IL-13 gene, its construction method and its application in the field of biomedicine. Background technique [0002] IL-13 was first discovered in 1989 and named P600, a protein preferentially produced by activated mouse Th2 cells. IL-13 belongs to type I cytokines. IL-13 has a strong immune regulation effect. These cell types have multiple functions, and these cell types are related to the pathogenesis of allergic diseases. IL-13 is an important mediator of allergic inflammation. In human B cells, human IL-13 and IL-4 have similar effects, Including promotion of B cell proliferation, induction of type conversion to IgG4 and IgE, induction of expression of surface antigens, including low-affinity IgE receptors CD23 (FcεRII) and MHC II, in monocytes and macrophages, and enhancement of many integrations b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027C12N15/24C12N5/10G01N33/68A61K49/00
CPCA01K67/0278A01K2207/15A01K2217/072A01K2227/105A01K2267/03A61K49/0008C07K14/5437C12N5/0606C12N15/8509C12N2510/00C12N2800/107G01N33/6854G01N33/6869G01N2333/5437
Inventor 沈月雷黄蕤白阳郭朝设张美玲尚诚彰
Owner BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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