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Application of iron transporter in treatment of Parkinson's disease

A technology for Parkinson's disease and disease, applied in the field of preparation of gene-targeted drugs

Active Publication Date: 2020-09-29
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some in vitro experiments show that iron ions may have a certain relationship with the pathogenesis of PD, there has been no more detailed and convincing evidence in vivo to prove

Method used

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  • Application of iron transporter in treatment of Parkinson's disease
  • Application of iron transporter in treatment of Parkinson's disease
  • Application of iron transporter in treatment of Parkinson's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1: Phylogenetic tree analysis

[0055] Using MEGA software to analyze the phylogenetic tree of human genes ZIP13 and Tsf, and Drosophila genes dZIP13 and Tsf1, it shows that Drosophila dZIP13 and human ZIP13, Drosophila Tsf1 and human Tsf belong to orthologous genes and have the same or similar functions.

Embodiment 2

[0056] Example 2: The acquisition of Tsf1 RNAi and dZIP13 OE transgenic Drosophila strains

[0057] 1. Using the genomic DNA of the wild-type Drosophila w1118 strain as a template, isolate and clone Tsf1 and dZIP13 by PCR, and then introduce them into Escherichia coli competent cells after digestion, ligation, and transformation. Positive clones were obtained through colony PCR, plasmid digestion identification, and sequencing identification.

[0058] 2. Using the wild-type Drosophila w1118 as the recipient, use microinjection technology to transform the constructed plasmid into newborn eggs through the method mediated by p-element. After the eggs develop into adults, number each Drosophila with w1118 After crossing, the red eyes of the offspring fruit flies indicate that the construction is successful.

[0059] Example 2: Effects of Drosophila dZIP13 and Tsf1 genes on PD-like behavior in PINK1-deficient Drosophila

[0060] 1. Effects of dZIP13 OE and Tsf1 RNAi on exercise c...

Embodiment 3

[0064] Example 3: Specific regulation of the expression levels of dZIP13 and Tsf1 in Drosophila muscle tissue can significantly affect mitochondrial respiration

[0065]In this experiment, Mhc-Gal4>+ / + normal group Drosophila, Mhc-Gal4>PINK1 RNAi model Drosophila, Mhc-Gal4>PINK1 RNAi; dZIP13 OE Drosophila and Mhc-Gal4>PINK1 RNAi; Tsf1 RNAi Drosophila trunk Separation is carried out, the supernatant is obtained by homogenization and centrifugation, and the ATP level of the supernatant is detected with a Promega ATP detection kit. Using seahorseXF-24 to detect the respiratory function of mitochondria in muscle, dissect the trunk of Drosophila, add mitochondria extract, use glass homogenizer to homogenize the tissue, and use gradient centrifugation to obtain mitochondrial pellet. The concentration of mitochondria was detected with a Bradford kit, and the mitochondria were added to seahorse 24-well plates. For the detection of mitochondrial complex III respiration, 4 mM sn-glycer...

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Abstract

The invention relates to the field of gene engineering, in particular to a target protein of Parkinson's disease (PD) and an encoding gene and application thereof, and particularly relates to preparation of a gene targeting drug aiming at the key gene. The target protein finds that a target gene ZIP13 and Transferrin (Tsf) for Parkinson's disease are provided, and the PD condition is remarkably relieved or delayed by over-expressing the ZIP13 or interfering the Tsf through gene silencing. According to the target protein the human PD target protein comprises amino acid sequences represented bySEQ ID No. 5 and SEQ ID No. 6, and the gene comprises nucleotide sequences represented by SEQ ID No. 7 and SEQ ID No. 8. According to the target gene, an RNAi vector carrying a target gene is constructed, and the athletic ability of a PD patient can be improved through gene silencing or gene overexpression, and PD symptoms caused by knockdown of PINK1 is relieved or prevented. The target protein is expected to provide a novel treatment strategy and a key target for treating hereditary PD.

Description

technical field [0001] The present invention relates to the field of genetic engineering. Specifically, the present invention relates to a target protein of Parkinson's Disease (PD) and its coding gene and application, especially to the preparation of gene-targeted drugs for the key genes mentioned above. Background technique [0002] Parkinson's Disease (PD) is a neurodegenerative disease that affects human health. The typical motor symptoms of PD have been considered an important part of the disease since its initial description by James Parkinson in the 19th century and later relabeled by Jean-Martin Charcot. These patients often manifest as muscle tremor, muscle rigidity, slow movement, dysgraphia, unsteady gait, general weakness, stiff facial expression, and depression, which makes it difficult for the patient to take care of himself, and brings heavy psychological and economic burdens to the family. affect social stability and harmony. At present, PD is considered to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/47C12N15/85A61K31/7105A61K48/00A61P25/16
CPCC07K14/47C12N15/85A61K31/7105A61K48/005A61P25/16
Inventor 肖桂然薛劲松韦田纪晓雯
Owner HEFEI UNIV OF TECH
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