Medulloblastoma animal model and establishment and application thereof

A technology of medulloblastoma and animal models, applied in the field of medulloblastoma animal models and its establishment and application, can solve the problem of lack of incidence of medulloblastoma

Active Publication Date: 2021-03-26
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is currently a lack of non-human mammalian models with a high incidence of medulloblastoma and genetically stable

Method used

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  • Medulloblastoma animal model and establishment and application thereof
  • Medulloblastoma animal model and establishment and application thereof
  • Medulloblastoma animal model and establishment and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0147] Embodiment 1 hybridization obtains Patched + / - / Trim32 KO transgenic mice

[0148] Patch first + / - Heterozygous male mice (Ptched + / - Mouse model (#003081) was purchased from JacksonLaboratory) with Trim32 KO Female mice are crossed to get Patched + / - / Trim32 + / - double heterozygous mice; then use double heterozygous mice with Trim32 KO Mice are backcrossed, and their offspring are selected as Patched + / - / Trim32 KO mouse, with Trim32 KO Two genotypes of mice have been continuously self-crossed for more than 3 generations to obtain Patched with stable genotypes + / - / Trim32 KO .

[0149] Patched + / - and Trim32 KO Mice Progeny mice were genotyped using the following primers, respectively.

[0150] Patched + / - Identification of mouse genotypes:

[0151] name sequence Numbering WT-f 5'-CTAGGCCACAGAATTGAAAGATCT-3' 1 WT-r 5'-GTAGGTGGAAATTCTAGCATCATCC-3' 2 Mut-f 5'-GATGATCTCGTCGTGACCCAT-3' 3 Mut-r 5'-GTAGGTACTCTGTTCT...

Embodiment 2

[0157] Example 2 Obtaining Math1-GFP / Dcx-DsRed / Patched by hybridization + / - / Trim32 KO transgenic mice

[0158] Math1-GFP transgenic mice were crossed with Dcx-DsRed transgenic mice to generate Math1-GFP / Dcx-DsRed transgenic mice.

[0159] Using Math1-GFP / Dcx-DsRed transgenic mice with Patched + / - Heterozygotes were crossed, and their progeny were continuously selfed for more than 3 generations to obtain Math1-GFP / Dcx-DsRed transgenic mice and Math1-GFP / Dcx-DsRed / Patched + / - transgenic mice.

[0160] Using Math1-GFP / Dcx-DsRed transgenic mice with Patched + / - / Trim32 KO Carry out hybridization, and take its progeny to continuously self-cross for more than 3 generations to obtain Math1-GFP / Dcx-DsRed / Patched + / - / Trim32 KO transgenic mice.

[0161] Math1-GFP and Dcx-DsRed transgenic mice were confirmed by green fluorescence or red fluorescence, while Patched + / - and Trim32 KO Mice Progeny mice were genotyped using the above primers, respectively.

[0162] result:

[0...

Embodiment 3

[0165] The establishment of embodiment 3 novel medulloblastoma model

[0166] 1. The incidence of MB

[0167] Obtain Patched by the method in embodiment 1 + / - / Trim32 KO Transgenic mice and Patched + / - / Trim32 WT , Trim32 KO , WT (ie Patched WT / Trim32 WT ) mice of four genotypes, the changes in biological behavior of each mouse were regularly observed during the period from 6 weeks to 40 weeks of age. severe weight loss, paralysis, or lack of activity), they were killed and autopsied. Histological analysis confirmed that the cerebellar tissue was medulloblastoma. Simultaneous statistical analysis of Ptch1 within 40 weeks + / - Incidence of MB in mice, and by Ptch1 + / - Progeny Ptch1 produced by crossing with Trim32 knockout mice + / - / Trim32 KO incidence of MB.

[0168] result:

[0169] like Figure 4 As shown, it was surprisingly found that knockout of Trim32 increased the incidence of MB from 25.4% to 52.9% in the background of Ptch1+ / - mice. 25.4% Ptch1 + / - / Tr...

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Abstract

The invention provides a medulloblastoma animal model and establishment and application thereof. The expression or activity of the Patched protein in the animal model cell is reduced, and the expression or activity of the Trim32 protein is reduced. The invention also provides a construction method of the non-human mammal model and a preparation method of the non-human mammal model of medulloblastoma carrying the cerebellar precursor cell marker and the early differentiated cell marker at the same time. The non-human mammal model is a more effective myeloblastoma (MB) model, the morbidity of MBis greatly improved compared with an existing classic model, and the non-human mammal model can be used as an excellent animal model for drug screening and experimental research.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an animal model of medulloblastoma and its establishment and application. Background technique [0002] Medulloblastoma occurs in the cerebellum and is one of the most common and malignant tumors of the nervous system. Children have a higher incidence, accounting for more than 70% of the total incidence of medulloblastoma. The incidence rate in children accounts for about 18.2% of brain tumors in children. Medulloblastoma tumors are fast growing, large in size, and rapidly progressive, resulting in a mortality rate nearly equal to the morbidity rate. [0003] The pathogenesis of medulloblastoma is still unclear, leading to difficulties in clinical treatment. It is of great scientific and social significance to develop new animal models of medulloblastoma and provide new ideas and means for elucidating its mechanism and developing new drugs. [0004] However, there is still a lack o...

Claims

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Application Information

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IPC IPC(8): A01K67/027C12N5/10C12Q1/02A61K45/00A61K38/17A61P35/00
CPCA01K67/0275C12N5/0692G01N33/5011A61K45/00A61K38/1709A61P35/00A01K2227/105A01K2267/0331C12N2510/00A61K2300/00
Inventor 高维强王明磊杨茹
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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