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

Construction method and application of transgenic rat specifically expressing hmrgprx4

A construction method, transgenic technology, applied in genetic engineering, biochemical equipment and methods, pharmaceutical formulations, etc., can solve problems such as genes without functional homology, and genetic methods cannot be used

Active Publication Date: 2022-04-29
HEPAITECH (BEIJING) BIOPHARMA TECH CO LTD
View PDF9 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, for the development of drugs for the treatment of human cholestatic pruritus, since MRGPRX4 is a primate-specific receptor, there is no functional homologous gene in both mice and rats 2 , so it is impossible to use traditional genetic methods, such as gene knockout, to study the function of this gene in model animals, and to screen candidate drugs in vivo

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 transgenic rat specifically expressing hmrgprx4
  • Construction method and application of transgenic rat specifically expressing hmrgprx4
  • Construction method and application of transgenic rat specifically expressing hmrgprx4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The present invention constructs a humanized rat by expressing human MRGPRX4 in rat itch sensory neurons, and provides an animal model for studying the function of MRGPRX4 and screening antipruritic drugs targeting human MRGPRX4. The specific method is as follows:

[0046] (1) Construction of hMRGPRX4-Cre rats

[0047] In the rat MrgA gene, hMRGPRX4-3xFlag-P2A-iCre-WPRE-pA (SEQ ID NO: 6) is used to replace the nucleotide sequence corresponding to the 28th amino acid on the MrgA gene exon 2 (exon2) to the terminator . The sgRNAs were designed in intron 1 (intron1) and exon 2 of the MrgA gene, respectively. Based on the design principle of sgRNA, multiple sgRNAs are designed in the target site area. The sgRNA (5′-TCCGGTTAGGAGTAAATTCC-3′ and 5′-TGAGACAGCGGAAAACACGG-3′) and Cas9 gene were connected to the plasmid vector pCS with T7 promoter and transcribed in vitro to obtain gRNA and Cas9 mRNA for microinjection.

[0048] According to the target gene sequence, the 5' an...

Embodiment 2

[0085] Figure 4 It further proved that the hMRGPRX4-Cre; lsl-hMRGPRX4-mScarlet transgenic rat has more advantages as an animal model. Such as Figure 4 As shown in a, hMRGPRX4-Cre homozygous rats were mated with a reporter rat, and under the action of Cre recombinase, a red fluorescent protein tdTomato would be expressed in the Cre-positive cells of the progeny rats. Extract and cultivate the DRG neurons of the offspring rats, such as Figure 4 As shown in b, the arrow shows the DRG neurons expressing tdTomato, indicating that the Cre recombinase is expressed and functional, and it also theoretically indicates that hMRGPRX4 is expressed; Figure 4 As shown in c, calcium imaging results showed that the addition of DCA did not cause obvious calcium signals in these neurons, indicating that although hMRGPRX4 was expressed, it may not mediate the response caused by DCA in DRG neurons due to the relatively low expression level. This also shows that it is necessary to increase t...

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 transgenic rat specifically expressing hMRGPRX4 and its application. The present invention successfully constructs a humanized rat by expressing hMRGPRX4 in rat itch neurons. The transgenic rat strains were mated to obtain transgenic rats specifically expressing hMRGPRX4. The invention provides an animal model for studying the function of hMRGPRX4 and screening antipruritic drugs targeting hMRGPRX4.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a transgenic rat specifically expressing hMRGPRX4 and its application. Background technique [0002] Patients with chronic liver disease are often accompanied by unbearable itching, and up to 60% of patients with primary biliary cholangitis (PBC) have symptoms of chronic itching, seriously affecting the quality of life of patients. However, there is still a lack of effective drugs for relieving or treating the pruritus. Recent studies have shown that human MRGPRX4 mediates bile acid-induced cholestatic pruritus. Cholic acid (CA) and its derivatives can activate hMRGPRX4; the content of cholic acid in the serum of chronic liver disease patients with pruritus is significantly higher than that of normal people; cholic acid and hMRGPRX4 agonists can induce obvious itching. Therefore, hMRGPRX4 is a drug target for the treatment of human cholestatic pruritus, a...

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): C12N15/85C12N15/65A01K67/027A61K49/00
CPCC12N15/8509C12N15/65A01K67/0278C07K14/723A61K49/0008C12N2800/107A01K2267/03A01K2267/0393A01K2227/105A01K2217/072A01K2207/15
Inventor 李毓龙赵天军雷晓光
Owner HEPAITECH (BEIJING) BIOPHARMA TECH CO LTD
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