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

Models of prion disease

a prion disease and model technology, applied in the field of system for studying neurodegenerative disorders, can solve the problems of long incubation period of prion when expressed in transgenic mice, limited immunological assay sensitivity, and in vitro methods for diagnosing prion disease in suspected diseased brains, such as detection of protease-resistant prp on western blots, and achieve the effect of rapid determination of the infectivity of prions in a sampl

Inactive Publication Date: 2005-03-03
RGT UNIV OF CALIFORNIA
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

Enables rapid infection and symptom development in transgenic animals, facilitating the study of prion disease progression and the identification of therapeutic agents, while providing a humane and cost-effective alternative to traditional animal models.

Problems solved by technology

Although previous Thr to Ala substitutions in the glycosylation sites of PrP resulted in conformational instability, the resulting PrPC molecules displayed an aberrant cellular localization and resulted in a very long prion incubation period when expressed in transgenic mice.
In vitro methods of diagnosing prion disease in a suspected diseased brain, such as detection of the presence of protease-resistant PrP on Western blots, is limited by the sensitivity of the immunological assay.

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

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0138] Mutations are introduced into human PrP sequences to alter two known glycosylation sites of the molecule. A mouse-hamster chimeric PrP transgene (MHM2) is mutagenized to contain a double mutation where the two glycosylation sites are substituted from asparagine to glutamine (N180Q / N196Q), and cloned into a pSPOX expression vector. The PrP molecule was mutagenized using a mismatched oligonucleotide primer and PCR amplification. Following amplification, the mutated PrP DNA is digested with unique restriction endonucleases to allow directional cloning into the pSPOX vector. These expression vectors were then transiently expressed in neuroblastoma cells (N2a cells) and subsequently infected with 10% brain homogenates from experimentally scrapie-infected mice.

[0139] After 4 days of incubation, appearance of newly formed prions can be seen after cell lysis, proteinase-digestion and immunoblot. The blot was developed using 3F4 (which sepcifically detects the MHM2 construct) as the ...

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 present invention provides a novel PrP protein, and nucleic acids encoding this protein, where the PrP protein is characterized in vivo by 1) incomplete glycosylation relative to glycosylation of wild-type PrPC and 2) proper cellular localization, i.e. an ability to be transported to the cell surface. This novel, under-glycosylated PrP, unlike its normal cellular counterpart, can easily be converted into a protease-resistant isoform by incubation with infectious prions. The invention further provides systems for the study of prion disorders and methods of using these systems, e.g. the study of the mechanical processes in progression of prion-mediated disease or the identification of new therapeutic agents for treatment of prion-mediated disorders. In such systems, protease-resistant under-glycosqvated PrP is generated de novo and can be detected by standard immunoblot techniques.

Description

GOVERNMENT RIGHTS [0001] The United States Government may have certain rights in this application pursuant to Grant No. AG10770 awarded by the National Institutes of Health.FIELD OF THE INVENTION [0002] The present invention relates generally to systems for studying neurodegenerative disorders, and in particular to systems for the study of prion-associated disease. BACKGROUND OF THE INVENTION [0003] Perons are infectious pathogens that cause central nervous system spongiform encephalopathies in humans and animals. -Prions are distinct from bacteria, viruses and viroids. The predominant hypothesis at present is that no nucleic acid component is necessary for infectivity of prion protein. Further, a prion which infects one species of animal (e.g., a human) will not readily infect another (e.g., a mouse). [0004] A major step in the study of prions and the diseases that they cause was the discovery and purification of a protein designated prion protein (“PrP”) [Bolton et al., Science 21...

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(United States)
IPC IPC(8): C07K14/47C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12N15/85C12Q1/02C12Q1/68A01K67/027G01N33/15G01N33/50G01N33/569G01N33/68
CPCA01K67/0275A01K2217/05A01K2227/105A01K2267/0318A01K2267/0337G01N2800/2828C07K14/47C12N15/8509G01N33/5088G01N33/6896G01N2500/00A01K2267/0343C07K14/00
Inventor PRUSINER, STANLEY B.KORTH, CARSTEN
Owner RGT UNIV OF CALIFORNIA
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