Nonsense mutation read-through activity use of blasticidin

A blasticidin and nonsense technology, applied in the field of biomedicine, can solve the problem of few types of nonsense mutation read-through agents

Inactive Publication Date: 2018-05-11
WUHAN UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention aims at the problem that there are few types of nonsense mutation read-throug

Method used

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  • Nonsense mutation read-through activity use of blasticidin
  • Nonsense mutation read-through activity use of blasticidin
  • Nonsense mutation read-through activity use of blasticidin

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Experimental program
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Embodiment Construction

[0017] The use of the nonsense mutation read-through activity of blasticidin will be further described below in conjunction with the experiments and results of extracellular and intracellular experiments.

[0018] 1) Detection of read-through activity of nonsense mutations of Blasticidin in luciferase cell model

[0019] We first constructed a lentiviral vector containing a nonsense mutation of the luciferase gene, packaged it into a lentivirus and infected the cells, established a stable cell model, and then treated the cells with blasticidin to read through the reported nonsense mutations The active drug G418 was used as a positive control, and the nonsense mutation read-through activity of blasticidin was identified by chemiluminescence. The following are the experimental data of the present invention:

[0020] (1) Construction of lentiviral vector containing nonsense mutation of luciferase gene

[0021] The cDNA plasmid pGL4-WT carrying wild-type luciferase 2 and the cDNA plasmid...

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Abstract

The invention discloses a nonsense mutation read-through activity function of blasticidin. A preliminary experiment result shows that the blasticidin can partially restore the expression of nonsense mutation-containing luciferase in a cell model, can partially restore the expression of a nonsense mutation-containing ATM gene in extracellular protein truncation experiments, is hopeful to develop new medicines a lead compound after being structurally reconstructed and modified to treat nonsense mutation induced various diseases, and has a potential medicine use.

Description

Technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to the use of nonsense mutation read-through activity of blasticidin. Background technique [0002] Among all human disease-causing gene mutations, 30% are nonsense mutations. Nonsense mutations are not only one of the main causes of genetic diseases, but also closely related to the onset of tumors. They have been found in more than 480 tumor-related genes. Nonsense mutations will form a stop code in the mRNA prematurely, leading to premature termination of mRNA translation at the mutation site, resulting in the loss of the encoded protein. Studies have found that a small number of compounds can induce the translation complex to pass the nonsense mutation site to read through, so that the mRNA of the mutant gene can continue to be translated, thereby restoring the expression of the functional full-length protein. [0003] A large number of studies have proved that for many diseases ...

Claims

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Application Information

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IPC IPC(8): A61K31/513A61P35/00A61P43/00
CPCA61K31/513
Inventor 江高峰袁辉杜柳涛
Owner WUHAN UNIV OF SCI & TECH
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