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

Construction method of PTDSS2 conditional gene knockout mouse model

A method for constructing a mouse model, applied in the field of genetic engineering, can solve the problem of not finding PTDSS2 gene conditional knockout mice, etc., and achieve the effect of low cost, time and cost saving

Active Publication Date: 2022-06-07
广东药康生物科技有限公司
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Currently, there are no relevant reports on the construction of PTDSS2 gene conditional knockout mice using CRISPR / Cas9 technology

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 PTDSS2 conditional gene knockout mouse model
  • Construction method of PTDSS2 conditional gene knockout mouse model
  • Construction method of PTDSS2 conditional gene knockout mouse model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1. Construction of PTDSS2 gene conditional knockout mouse model

[0039] This example uses CRISPR Cas9 technology to genetically modify the mouse PTDSS2 gene. After the mice with stable genetic inheritance are mated with mice expressing Cre recombinase, the flox mice are knocked out, and knocking out this region will lead to the destruction of the protein function . Unless otherwise specified, the mice used in this example are C57BL / 6J strain mice.

[0040] 1. Determine the conditional knockout region and sequence

[0041] According to the structure of the PTDSS2 gene, Exon2-Exon3 of the PTDSS2-201 transcript was used as the knockout region, which contained a 185 bp coding sequence. The sequence of the targeting vector is shown below, where capital letters underlined correspond to Loxp sites.

[0042] The insertion position of Loxp1 is as follows:

[0043] cttgaagacctgacctgggaccggccctgtccctttcttggcaacgccttcatggttccctggttggttctggcagggttgccttcctacccactcagcgatt...

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 method for constructing a PTDSS2 gene conditional knockout mouse model by using a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 (CRISPR associated protein 9) technology. The method comprises the following steps: 1, designing sgRNA (small guide ribonucleic acid) aiming at mouse source PTDSS2 gene Exon2-Exon3, 2, obtaining the sgRNA by using an in-vitro transcription technology, and 3, co-injecting or co-electrically transferring a targeting vector, the sgRNA and Cas9 protein to a mouse fertilized egg. Compared with a traditional ES targeting mouse model, the PTDSS2 gene conditional knockout mouse model constructed for the first time has the characteristics of high efficiency, rapidness, simplicity, convenience, low cost and the like, and the gene can be regulated and controlled to be expressed in specific tissues or specific time through Cre.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for constructing a PTDSS2 conditional gene knockout mouse model and its application. Background technique [0002] CRISPR / Cas9 technology realizes specific DNA recognition by designing specific sgRNA, and completes cutting at the target position, and then completes the repair of the break through the DNA repair mechanism of the cell itself, thereby realizing the "editing" of the target gene. With the development and in-depth research of CRISPR / Cas9 technology, it has been widely used to construct transgenic model animals (mainly gene-edited mice), thus providing basic research on animal-level treatments for preclinical use. [0003] The pathogenesis of human diseases and the screening of effective therapeutic drugs require extensive preclinical trials. Due to ethical restrictions on the direct use of human cells and tissues for preclinical research, animal...

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): C12N15/89C12N15/87C12N9/22C12N15/113C12N15/54A01K67/027C12Q1/6888C12N15/11A61K49/00
CPCC12N15/89C12N15/87C12N9/22C12N15/1137C12N9/1288A01K67/0276C12Q1/6888A61K49/0008C12N2310/20C12Y207/08029A01K2217/075A01K2217/15A01K2217/206A01K2227/105A01K2267/0306C12Q2600/156C12Q2600/124
Inventor 李颖王韬王宏宇蒋余亭陈景曦黎晓雯郑桂纯
Owner 广东药康生物科技有限公司
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