Unlock instant, AI-driven research and patent intelligence for your innovation.

Construction method and application of humanized ccr2 gene modified animal model

A technology of genetic modification, construction method, applied in the field of biology

Active Publication Date: 2022-05-27
SHANGHAI BIOMODEL ORGANISM SCI & TECH DEV
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The screening and preclinical tests of human CCR2 target drugs need to be evaluated in animal models, but due to the difference in species, the homology between human CCR2 and mouse Ccr2 protein is only 68%, and the ones screened in mice Targeted drugs are not fully applicable to humans, and drugs targeting human CCR2 protein may not necessarily recognize mouse Ccr2 protein

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 and application of humanized ccr2 gene modified animal model
  • Construction method and application of humanized ccr2 gene modified animal model
  • Construction method and application of humanized ccr2 gene modified animal model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Sequence design

[0065] Both the mouse Ccr2 gene and the human CCR2 gene contain multiple transcripts, and the sequence design in this example is mainly described by taking one of the transcripts as an example. That is, the main part of exon 3 of the mouse Ccr2 gene (NCBI Gene ID: 12772) (based on the transcript of the NCBI accession number NM_009915.2→NP_034045.1, the mRNA sequence of which is shown in SEQ ID NO: 1, The corresponding protein sequence is shown in SEQ ID NO: 2) is replaced with the corresponding fragment of the human CCR2 gene (Gene ID: 729230) (based on the transcript of NCBI accession number NM_001123396.3→NP_001116868.1, its mRNA sequence is as SEQ ID NO: 3, the corresponding protein sequence is shown in SEQ ID NO: 4), wherein, the comparison diagram of mouse Ccr2 and human CCR2 gene is shown in figure 1 , see the schematic diagram of the final modified humanized mouse CCR2 gene figure 2 , the humanized mouse CCR2 gene DNA sequence (chim...

Embodiment 2

[0071] Example 2 Design and construction of recombinant vector PBR322-CCR2

[0072] According to the sequence design, the inventors further designed such as image 3 Targeting scheme shown and vector containing 5' homology arm, human CCR2 gene fragment, 3' homology arm. Wherein the 5' homology arm (SEQ ID NO: 9) is the 124102766-124105684 nucleotide of the NCBI accession number NC_000075.6, and the 3' homology arm (SEQ ID NO: 10) is the NCBI accession number NC_000075. The 124106806-124110723 nucleotides of 6, the human CCR2 (SEQ ID NO: 11) gene fragment is the 46357528-46358610 nucleotides of the NCBI accession number NC_000003.12.

[0073] The construction process of the vector is as follows: upstream primers for amplifying three homologous recombination fragments (LA, KI, RA) and matching downstream primers and related sequences are designed. Among them, the 5' homology arm corresponds to the LA fragment, the human CCR2 gene fragment corresponds to the KI fragment, and th...

Embodiment 3

[0084] Example 3 Validation of the vector PBR322-CCR2

[0085] Five PBR322-CCR2 clones were randomly selected and verified by restriction endonuclease EcoRI. The electrophoresis of the digestion products should show a fragment of size 5614bp+4433bp+2759bp+1038bp. For enzyme digestion results, see Figure 4 , the results of plasmid digestion were in line with expectations, indicating that the results of plasmid digestion verification were correct. The plasmid was sequenced by a sequencing company and verified to be correct, and follow-up experiments were carried out.

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 provides a method for constructing a humanized CCR2 genetically modified animal model and its application, and relates to the field of biotechnology. The humanized CCR2 genetically modified animal model constructed by the present invention can accelerate the research progress in the fields related to human CCR2 gene or protein. Ex-experiments provide valid models and powerful tools. Preferably, the present invention uses CRISPR / Cas9 gene editing technology to replace the mouse Ccr2 gene with the human CCR2 gene to construct a mouse model that can interact with the anti-human CCR2 antibody. Compared with ordinary mice, the model has The humanized transformation of key target molecules has been realized, which can be used to screen and evaluate drugs targeting human CCR2 gene, and is an ideal model for preclinical drug testing.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a humanized CCR2 gene-modified animal model and its application. Background technique [0002] Laboratory animal disease models are indispensable research tools for studying the etiology and pathogenesis of human diseases, and for developing prevention and treatment technologies and drugs. Common experimental animals include mice, rats, guinea pigs, hamsters (hamsters), rabbits, dogs, monkeys, pigs, fish, etc. Although most human genes have homologous genes in animals, there are still many differences between humans and animals in gene sequences and protein sequences, resulting in many drugs targeting human targets cannot be combined with homologous proteins in animals. Pharmacological efficacy, which is particularly significant in macromolecular antibody drugs, because macromolecular antibody drugs are highly specific, and a single amino acid difference ma...

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 Patents(China)
IPC IPC(8): C12N15/90C12N15/85C12N15/877C12N5/10C12N5/071C12N15/12A01K67/027A61K49/00
CPCC12N15/907C12N15/8509C12N15/8775C07K14/723A01K67/0278C12N5/0604C12N5/0602A61K49/0008C12N2800/107A01K2217/072A01K2227/105A01K2267/0331C12N2510/00C12N2503/02C12N2517/02
Inventor 孙瑞林王津津慈磊
Owner SHANGHAI BIOMODEL ORGANISM SCI & TECH DEV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com