Variant p53 gene and recombinant adenovirus for cancer treatment

A p53 gene, recombinant adenovirus technology, applied in the field of genetic engineering, can solve problems such as protein failure, influence, and protein expression level.

Pending Publication Date: 2020-06-23
SYNO SHENZHEN BIOMEDICAL RES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In most cases, the cause of a gene defect is likely to be due to a mutation that affects its protein structure and forces it not to function properly, or a mutation that affects the expression level of the protein leading to a complete failure of the protein to highlight or improper expression during the cell cycle

Method used

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  • Variant p53 gene and recombinant adenovirus for cancer treatment
  • Variant p53 gene and recombinant adenovirus for cancer treatment
  • Variant p53 gene and recombinant adenovirus for cancer treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0032] This example will detail the construction of a recombinant adenovirus containing a variant p53 gene.

[0033] This example uses the original gene therapy virus vector rAd-RSV-p53 (recombinant human p53 adenovirus produced by Shenzhen Saibainuo Gene Technology Co., Ltd., trade name Gendicine) as the raw material, and the vector can be purchased from market channels.

[0034] The length of the genomic DNA in the viral vector rAd-RSV-p53 is 35698bp, the data comes from the whole genome structure sequence of human adenovirus type 5 (refer to GenBank NC_001406), and the results of vector Sanger sequencing, such as figure 1shown in . The Sanger sequencing results of the p53 expression cassette are shown in SeQ ID No.1 in the sequence listing.

[0035] figure 1 is the linear DNA structure diagram of the viral vector rAd-RSV-p53, which shows the position of the p53 expression cassette, and the part from the left end to the first SalI restriction site.

[0036] The first thin...

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PUM

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Abstract

The invention discloses a variant p53 gene for cancer treatment. A cDNA base sequence of the p53 gene is constructed by replacing codon 72 in a cDNA base sequence of a naturally occurring p53 gene with a target codon. The invention also discloses a recombinant adenovirus containing the variant p53 gene. The recombinant adenovirus is constructed by artificially synthesizing a DNA target fragment containing the target codon, replacing the DNA target fragment with a DNA fragment containing a p53 expression cassette in an intermediate plasmid to generate a variant intermediate plasmid containing the variant p53 gene, and introducing the DNA fragment containing the variant p53 gene in the variant intermediate plasmid into a shuttle plasmid, and then performing homologous recombination to obtainthe recombinant adenovirus containing the variant p53 gene.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a series of variant p53 genes used for gene enhancement, addition and complementary cancer treatment, and a construction method of a recombinant adenovirus containing the variant p53 gene. Background technique [0002] The use of genes to treat various human diseases has been envisioned since it was established that defective gene function is a major cause of genetic diseases. In most cases, the cause of a gene defect is likely to be due to a mutation that affects its protein structure and forces it not to function properly, or a mutation that affects the expression level of the protein leading to a complete failure of the protein to highlight or improper expression during the cell cycle . Therefore, the goal of gene therapy is to introduce therapeutic genes into target cells and repair those disease-inducing genes, so as to restore the normal function of target cells ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/861A61K48/00A61P35/00
CPCC07K14/4746C12N15/86A61K48/005A61P35/00C12N2710/10043
Inventor 迪奥斯达德·鲍蒂斯塔徐卫
Owner SYNO SHENZHEN BIOMEDICAL RES CO LTD
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