Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Construction method of AURKA-CKO1-N condition gene knockout mouse model

A technology for gene knockout mice and construction methods, which can be used in genetic engineering, plant genetic improvement, botany equipment and methods, etc., and can solve the problems of mouse embryo lethality, etc.

Active Publication Date: 2019-01-15
XUZHOU MEDICAL UNIV
View PDF9 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanism of Aurora-A involved in tumorigenesis and development needs to be further elucidated
However, since Aurora-A systemic knockout mice are prone to mouse embryonic lethality

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Construction method of AURKA-CKO1-N condition gene knockout mouse model
  • Construction method of AURKA-CKO1-N condition gene knockout mouse model
  • Construction method of AURKA-CKO1-N condition gene knockout mouse model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] This embodiment provides a method for constructing an AURKA-CKO1-N conditional gene knockout mouse model, comprising the following steps:

[0047] (1) Construction of a recombinant targeting vector for AURKA conditional gene knockout

[0048]Use the ES cell (JM8A3ES cell, derived from C57 / BL6N strain) vector to target the AURKA gene, and use the bacterial artificial chromosome (BAC) for targeting, so that two loxP sites are inserted into intron 2 and intron 3 respectively. When Cre is expressed When exon 3 of the mouse AURKA gene can be knocked out, and replaced with the Neo gene, resulting in a frameshift mutation, the protein translation is terminated in exon 3 in advance;

[0049] The upstream arm is 4.2kb (DNA kilobase pairs), the downstream arm is 4.0kb, and the downstream arm is flanked by the phosphoglycerate kinase promoter-mediated negative selection marker gene expression cassette of herpes simplex virus thymidine kinase, and finally the AURKA conditional Gen...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a construction method of an AURKA-CKO1-N condition gene knockout mouse model. A recombination targeting carrier is constructed, embryonic stem cell are transfected after linearization, embryonic stem cell with homologous recombination are screened for clone, a donor mice blastula is injected after amplification to produce a chimera mouse; the chimera mouse copulates with an flp mouse to obtain a male F1 generation of mouse; and the male F1 generation of mouse copulates with a mouse whose tissues include specific Cre recombinase, and the AURKA-CKO1-N condition gene knockout mouse model is obtained in the next generation. According to the AURKA-CKO1-N condition gene knockout mouse model, an AURKA gene loses functions in specific tissues and cells, and an ideal modelis provided for research on an action mechanism of Aurora-A in tumorigenesis development.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for constructing an AURKA-CKO1-N conditional gene knockout mouse model. Background technique [0002] Gene knockout is a new technology developed in the second half of the 1980s by applying the principle of DNA homologous recombination. The definition of gene knockout refers to a gene whose structure is known but whose function is unknown, an experiment is designed at the molecular level to remove the gene, and then the experimental animal is observed as a whole to speculate on the function of the corresponding gene. [0003] Aurora-A is an important class of serine / threonine protein kinase involved in the regulation of cell mitosis, and its coding gene is located in the 20q13.2 chromosome region where translocation, deletion and amplification are active. Means that its expression pattern is inherently unstable. Abnormal expression of Aurora-A can le...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027C12N15/54C12N15/85
CPCA01K67/0276A01K2217/075A01K2227/105C12N9/12C12Y207/11001
Inventor 杨晶郑葵阳汤仁仙
Owner XUZHOU MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com