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Chimera for specifically degrading tau protein, encoding gene of chimera and application of chimera and encoding gene

A chimera and protein technology, applied in the field of genetic engineering, can solve serious side effects and other problems, and achieve the effect of high potency and reduced content

Pending Publication Date: 2020-08-11
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

The inventors discovered another type of small molecule called TAU which was found to break down certain proteins involved with neurology disorders like Parkinson’s Disease. They were able to clone this tiny chemical from bacteria or other organisms for use on humans without causing any harmful side effect(like create new ones). These smaller compounds could potentially help stop these brain conditions caused by specific enzymes such as beta amylase.

Problems solved by technology

This patented technical problem addressed in this patents is how effective therapies aimed at decreasing cognitive declination related disorders like Alzheimers disease (AD)/tau-associated amnesiodevelopments (TMAs)-affected memory impairment (TAFI)). Current methods have limitations due to their lack of specificity and possible negative impacts on brain function.

Method used

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  • Chimera for specifically degrading tau protein, encoding gene of chimera and application of chimera and encoding gene
  • Chimera for specifically degrading tau protein, encoding gene of chimera and application of chimera and encoding gene
  • Chimera for specifically degrading tau protein, encoding gene of chimera and application of chimera and encoding gene

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Embodiment approach

[0046] According to a preferred embodiment of the present invention, the vector is an adenovirus vector, and an adenovirus is a macromolecule (36 kb) double-stranded non-enveloped DNA virus, which enters the cell through receptor-mediated endocytosis, Then the adenovirus genome is transferred to the nucleus, remains outside the chromosome, and does not integrate into the host cell genome. Therefore, by integrating the target gene into the adenovirus vector, the target gene can be effectively delivered into the cell without affecting the cell's chromosome. Further preferably, the adenovirus vector is an adenovirus AOV002 vector.

[0047] Wherein, the target gene can be inserted into the conventional insertion site of the adenovirus vector, preferably, the target gene is inserted into the reading frame between the EcoRI and BamH I restriction sites of the adenovirus vector. Specifically, the adeno-associated virus (Adeno-associated virus (Adeno- associated virus, AAV), referre...

preparation example

[0057] Entrust Heyuan Biotechnology (Shanghai) Co., Ltd. to complete the construction and packaging of the following vectors.

[0058] (1) The pIRES2-EGFP plasmid expressing the chimera (β-tubulin-XIAP, SEQ.ID.NO:4) of the present application (the expression cassette is CMV-β-tubulin-XIAP-FLAG-IRES2-EGFP).

[0059] (2) Recombinant adeno-associated virus AAV-PROTAC (pAAV-CAG-β-tubulin-XIAP-2A-EGFP-3FLAG, recombinant adeno The titer of the virus reached 4.00×10 12 More than μg / mL indicates a higher titer.

[0060] (3) Adeno-associated virus AAV-hTau-Flag (AAV-hSyn-hTau-Flag) used to inject wild mice to express human tau protein.

[0061] (4) Control adeno-associated virus AAV-Flag.

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Abstract

The invention relates to the technical field of gene engineering, in particular to a chimera for specifically degrading tau protein, an encoding gene of the chimera and application of the chimera andthe encoding gene. The chimera comprises a tau protein binding sequence, a connecting sequence and a ubiquitin ligase recruiting sequence, wherein the tau protein binding sequence is connected with the ubiquitin ligase recruiting sequence through the connecting sequence. The chimeric capable of specifically degrading the tau protein can enhance the degradation of the tau protein in cells, so thatthe content of the tau protein is reduced; a chimera in the AAV-PROTAC can be normally expressed in a hippocampal region of a C57BL/6 mouse, and human tau protein is effectively degraded; AAV-PROTAC is injected into the hippocampal region of a 3xTg AD model mouse, so that the human tau protein can be effectively degraded. Therefore, the chimera capable of specifically degrading the tau protein canplay a role in preventing and treating a series of tau diseases including Alzheimer's disease.

Description

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Claims

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Application Information

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Owner HUAZHONG UNIV OF SCI & TECH
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