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Polypeptide derived from RPS23RG1 and application of polypeptide

A derivative peptide, selected technology, applied in the field of molecular biology and disease treatment, disease treatment, can solve problems such as difficult to develop drugs for the treatment of neurodegenerative diseases

Active Publication Date: 2019-04-09
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the specific molecular mechanism of RPS23RG1 protein in AD is still unknown, so it is difficult to develop drugs based on RPS23RG1 protein that can be effectively used in the treatment of neurodegenerative diseases

Method used

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  • Polypeptide derived from RPS23RG1 and application of polypeptide
  • Polypeptide derived from RPS23RG1 and application of polypeptide
  • Polypeptide derived from RPS23RG1 and application of polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0378] Example 1: Construction and Characteristic Analysis of Rps23rg1 Knockout Mice

[0379] 1.1 Construction of gene knockout mice

[0380] TALEN (transcription activator-like effector nucleases) recognizes DNA sequences through transcription activator-like effectors (TALEs), and cuts DNA at specific sites, forming double-strand breaks (DSBs), followed by non-homologous end joining (NHEJ ) repair mechanism to form random insertion or deletion of multiple bases, thereby realizing gene knockout. In this example, Rps23rg1 gene knockout mice were constructed using TALEN gene targeting technology.

[0381] First, according to the Rps23rg1 gene sequence on NCBI (GenBank ID: 546049), two TALEN targeting binding sequences were designed on exon 6 of the Rps23rg1 gene: TAL EN-L (targeting 5'-TGACCTTTTTCGACGAAATCCAG-3'; SEQ ID NO: 60) and TALEN-R (targeting 5'-TGCCTGAACTTCATTGAGG AAA-3'; SEQ ID NO: 61), the TALEN targeting schematic diagram is as follows figure 1 As shown, the mouse...

Embodiment 2

[0389] Example 2: Expression of PSD-95 and PSD-93 in Rps23rg1 knockout mice

[0390] First, the researchers of the present invention detected the expression of synapse-related proteins in Rps23rg1 knockout mice, and the results were as follows: Figures 5A-5C shown. Figure 5A The results of Western blot detection of the expression of synapse-related proteins in the brain tissue of mice are shown, and the results show that the protein expression levels of PSD-95 and PSD-93 in the brain tissue of KO mice are significantly lower than those of WT mice. Figure 5B The results of Western blot detection of protein expression in synaptosomes of mice are shown, and the results show that the protein expression levels of PSD-95 and PSD-93 in synaptosomes of KO mice are significantly lower than those of WT mice. Figure 5C The results of immunofluorescent staining of primary neurons from WT and KO mice are shown, and the results showed that PSD-95 and PSD-93 were significantly reduced i...

Embodiment 3

[0393] Example 3: Interaction of RPS23RG1 with PSD-95 and PSD-93 and its Effect on PSD-95 and PSD-93 Degradation

[0394] RPS23RG1 is a typical Ib transmembrane protein. Based on the experimental results obtained in Example 2, the intracellular segment of RPS23RG1 may have the potential to interact with PSD-95 and PSD-93 to affect the ubiquitination of PSD-95 and PSD-93 . Therefore, the researchers of the present invention analyzed the interaction of RPS23RG1 with PSD-95 and PSD-93 by immunoprecipitation-immunoblotting.

[0395] First, the researchers of the present invention constructed HA-modified mouse-derived full-length RPS23RG1 (1-141) and different truncations (1-134, 1-130, 1-116), see Table 2 and Figure 8A . The nucleic acid sequences encoding the above polypeptides were respectively inserted into pCMVHA plasmids to obtain plasmids expressing the above polypeptides. The above plasmids were transfected into HEK 293T cells to overexpress full-length and different tr...

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PUM

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Abstract

The invention relates to the fields of molecular biologics and disease treatment, in particular to the field of treatment on diseases related to too low activity of PSD-95 and / or PSD-93. Specifically,the invention relates to polypeptide (or variants thereof) for treating diseases related to too low activity of PSD-95 and / or PSD-93 (such as nervous system diseases), a fusion protein with the polypeptide (or variants thereof), and application of the polypeptide (or variants thereof) and the fusion protein. The invention further relates to a medicine composition for treating or alleviating one or more symptoms of diseases related to too low activity of PSD-95 and / or PSD-93 (such as nervous system diseases), wherein the medicine composition comprises the polypeptide (or variants thereof) or the fusion protein.

Description

technical field [0001] The invention relates to the field of molecular biology and disease treatment, especially the field of disease treatment related to the low activity of PSD-95 and / or PSD-93. In particular, the present invention relates to polypeptides (or variants thereof) useful for treating diseases associated with hypoactivity of PSD-95 and / or PSD-93 (for example, neurological diseases), comprising such polypeptides (or variants thereof) body), and uses of such polypeptides (or variants thereof) and fusion proteins. The invention also relates to pharmaceutical compositions useful for treating or alleviating one or more symptoms of diseases associated with hypoactivity of PSD-95 and / or PSD-93 (e.g., neurological diseases), comprising a polypeptide of the invention (or its variants) or fusion protein. Background technique [0002] Alzheimer's disease (AD), also known as senile dementia, is the most common cognitive impairment neurodegenerative disease closely relate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K19/00C12N15/12C12N15/62A61K38/17A61K47/42A61P25/28A61P25/14A61P25/00
CPCA61K38/00A61P25/00A61P25/14A61P25/28C07K14/4711C07K2319/00C07K2319/10C07K2319/33A61K38/17A61K47/42C07K14/435C07K14/47C07K19/00C12N15/62C12N15/79
Inventor 张云武赵东栋张慕娴孟健许华曦
Owner XIAMEN UNIV
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