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CRTAP gene and expression product thereof capable of serving as target for diagnosing and treating Alzheimer's disease

A technology for Alzheimer's disease and expression products, which can be used in gene therapy, microbial determination/examination, DNA/RNA fragments, etc., and can solve problems such as unclear etiology and pathogenesis, and inability to tissue reversal of the disease.

Active Publication Date: 2015-11-04
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since the etiology and pathogenesis of AD are still unclear, there is currently no specific method to reverse and prevent the progression of the disease. Therefore, at present, symptomatic treatment is mainly carried out at an early stage, and currently clinically diagnosed AD patients are basically in the middle and late stages, and existing treatments can only improve Symptoms, do not prevent or reverse disease progression

Method used

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  • CRTAP gene and expression product thereof capable of serving as target for diagnosing and treating Alzheimer's disease
  • CRTAP gene and expression product thereof capable of serving as target for diagnosing and treating Alzheimer's disease
  • CRTAP gene and expression product thereof capable of serving as target for diagnosing and treating Alzheimer's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Screening for Gene Markers Related to Alzheimer's Disease

[0057] 1. Collection of peripheral blood samples

[0058] AD patients came from Beijing 301 Hospital, a total of 60 cases, aged 53-84 years old, all cases were diagnosed as AD, and its diagnostic criteria refer to the third revised edition of the American Diagnostic and Statistical Manual of Mental Disorders. A total of 50 cases were selected as the control group, selected from the routine physical examination population of Beijing Hospital. All the subjects were excluded from blood lipid metabolism and other diseases, aged 60-82 years. All subjects signed the informed consent for the testing project and provided peripheral blood for genetic testing.

[0059] 2. Total RNA extraction from blood

[0060] Blood total RNA was extracted using Biotech Blood RNA Extraction Kit.

[0061] (1) Take 250 μl (or 0.25 g) of whole blood into an RNase-Free filter column, centrifuge at 13,000 rpm for 2 minutes, col...

Embodiment 2

[0091] Embodiment 2 interferes with the expression of CRTAP gene

[0092] 1. siRNA design and synthesis

[0093] siRNA sequences against CRTAP:

[0094] siRNA1-CRTAP:

[0095] The sense strand is 5'-UUAUUUGCCUUGAAGUAAGCG-3' (SEQ ID NO.5);

[0096] The antisense strand is 5'-CUUACUUCAAGGCAAAUAAUC-3' (SEQ ID NO.6),

[0097] siRNA2-CRTAP:

[0098] The sense strand is 5'-UAUGGAAAGGUAGAAAUCCUU-3' (SEQ ID NO.7);

[0099] The antisense strand is 5'-GGAUUUCUACCUUUCCAUAGC-3' (SEQ ID NO.8),

[0100] siRNA3-CRTAP:

[0101] The sense strand is 5'-UCGUUCAACUUAUAAUAGGCA-3' (SEQ ID NO.9);

[0102] The antisense strand is 5'-CCUAUUAUAAGUUGAACGACC-3' (SEQ ID NO.10)

[0103] Negative control siRNA sequence (siRNA-NC):

[0104] The sense strand is 5'-CGUACGCGGAAUACUUCGA-3' (SEQ ID NO.13);

[0105] The antisense strand is 5'-UCGAAGUAUUCCGCGUACG-3' (SEQ ID NO.14).

[0106] Neural cell line R2L1 cells were divided into 1×10 4 Inoculate / well into 24-well cell culture plates at 37°C, 5% C...

Embodiment 3

[0116] Example 3 Antagonistic effect of CRTAP gene on neuronal cell death induced by Aβ

[0117] 1. Cell transfection: according to the method in Example 2, the neural cell line R2L1 was transfected with siRNA1-CRTAP and siRNA-NC.

[0118] 2. After 24 hours of transfection, the neural cell line R2L1 was cultured in a 96-well plate with a cell density of 0.5×10 4 cells / well. Divide the cells into the following groups:

[0119] Uninduced group: transfected with siRNA-NC, without adding 20 μM Aβ42;

[0120] Negative control group (siRNA-NC+Aβ42): siRNA-NC was transfected, and 20 μM Aβ42 was added to the medium;

[0121] CRTAP gene interference group (siRNA-CRTAP+Aβ42): siRNA-CRTAP was transfected, and 20 μM Aβ42 was added to the medium;

[0122] Three replicate holes were set up for each group. After incubation at 37° C. for 20 h, MTT was added and incubated for 4 h. After adding the dissolving solution, incubate at 37°C for another 12h, and read the optical density value a...

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Abstract

The invention discloses a CRTAP gene and an expression product thereof capable of serving as a molecular marker for early diagnosis of Alzheimer's disease, namely judging whether a subject suffers from the Alzheimer's disease by detecting the CRTAP gene expression level in blood of the subject. According to the research result of the invention, a medicament capable of inhibiting CRTAP gene expression or inhibiting functions of a CRTAP gene expression product can be researched, thus realizing prevention and treatment of the Alzheimer's disease in clinic.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of human CRTAP gene in the diagnosis and treatment of Alzheimer's disease. Background technique [0002] Alzheimer's disease (Alzheimer disease, AD), also known as senile dementia, is a degenerative disease of the central nervous system with an insidious onset and a chronic progressive course. It is the most common type of senile dementia. The main manifestations are neuropsychiatric symptoms such as progressive memory impairment, cognitive dysfunction, personality changes, and language barriers, which seriously affect social, occupational, and life functions. The etiology and pathogenesis of AD have not yet been elucidated, and the characteristic pathological changes are extracellular senile plaques formed by the deposition of β-amyloid protein, neurofibrillary tangles formed by hyperphosphorylation of tau protein, and neuron loss with glial cell proliferation Wait. [000...

Claims

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Application Information

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IPC IPC(8): C12N15/11C07K16/18C12Q1/68G01N33/53C40B40/08C40B40/10A61K48/00A61P25/28
Inventor 杨承刚李曙光
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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