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Model for neurodegenerative disease

A technology of transgenic animals and animals, applied in the field of neurodegenerative diseases, can solve the problem of knowing downstream intracellular effectors

Inactive Publication Date: 2007-12-05
CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS (CSIC) +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Although activation of glycogen synthase kinase-3β (GSK-3β) has been suggested to be responsible for neuronal dysfunction, little is known about the downstream intracellular effectors responsible for this neuronal dysfunction

Method used

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Embodiment approach

[0022] We preferentially used the tet-regulatory system in mice. This tet-regulatory system has been used for conditional gene expression in eukaryotic cell systems and in mice (Gingrich and Roder, 1998). By using this system to prime the transgenic expression of mutant types of huntingtin, some of us have recently established the first conditional animal models of neurodegenerative diseases (Yamamoto et al., 2000). The tet-regulatory system is particularly useful when mimicking pathological conditions, as it can be used to: prevent perinatal lethality due to transgene toxicity, initiate transgene expression only in adulthood, and once the associated phenotype changes That is, transgene expression is stopped (Kelz et al., 1999; Yamamoto et al., 2000).

[0023] Regulation of this system is achieved by tetracycline-regulated transactivator (tTA), a chimeric protein consisting of a tet-repressor DNA-binding domain and a VP16 transactivation domain ( Gossen and Bujard, 1992). T...

Embodiment

[0026] The present invention is further illustrated by the following examples of our experimental work. Generation of injection fragments

[0027]The 8.0 kb Ase I fragment (BitetO) was used for microinjection. To generate BitetO, the 1.5 kb Hind III fragment corresponding to the Xenopus GSK-3β cDNA with the N-terminal MYC epitope was excised from the pcDNA3-GSK3 plasmid (Sanchez et al., 2000). This fragment was subcloned into the pCRII cloning vector (Invitrogen) digested with Hind III. Digest with XhoI to test for correct orientation. Then, by Nsi I-Not I digestion, a 1.5 kb fragment was excised and subcloned into the Pst I-Not I site of a plasmid containing a bidirectional tetO sequence that was replaced by a giant lacZ reporter gene sequence. Cytovirus (CMV) minimal promoter flanking (pBI-3, (Baron et al., 1995)). Finally, the 8.0 kb Ase I BitetO fragment was microinjected into single-cell CBAxC57BL / 6 embryos. Founder mice were identified using PCR and confirmed by Sou...

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Abstract

A transgenic animal in which GSK-3beta protein is over-expressed is useful as a model for neurodegenerative disease.

Description

[0001] The present invention relates to animal models for neurodegenerative diseases, in particular Alzheimer's disease. Background of the Invention [0002] Alzheimer's disease (AD), one of the most common neurodegenerative diseases in developed countries, is characterized by progressive memory loss and impairment of language and behavior, ultimately leading to death (Alzheimer, 1911; Yankner, 1996 ). Cognitive decline in AD is accompanied by neuronal atrophy and loss primarily in the cortex, hippocampus and tonsils (Gomez-Isla et al., 1997). In addition to a specific pattern of neuronal cell death, AD is also characterized by two neuropathological hallmarks: senile plaques and neurofibrillary tangles (NFTs). [0003] Senile plaques are extracellular deposits of amyloid fibrils composed of 39-43 amino acid beta-amyloid peptides (ABs) that are usually surrounded by dystrophic axons (Glenner and Wong, 2007). 1984; Masters et al., 1985; Selkoe, 1994). [0004] NFTs are collec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/56A61K49/00A01K67/027C12N9/12C12N15/09C12N15/85
CPCC12N9/1205A01K2227/105A01K2217/20A01K2267/0318A01K2217/05A01K2267/0312A01K67/0275C12N15/8509A01K2217/00A01K2207/15A01K67/027
Inventor 费利克斯·埃尔南德斯·佩雷斯赫苏斯·阿维拉·德格拉多何塞·哈维尔·卢卡斯·洛萨诺
Owner CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS (CSIC)
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