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pdl-1 gene target-specific methylation epigenetic modification anti-tumor molecule and use thereof

A PD-L1, specific technology, applied in the direction of anti-tumor drugs, genetic engineering, plant gene improvement, etc., can solve the problems of anti-tumor effect limitation

Active Publication Date: 2021-06-04
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has now been confirmed that the overexpression of PD-L1 on the surface of tumor cells significantly limits the antitumor effects of macrophages, DCs, MDSCs, etc.

Method used

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  • pdl-1 gene target-specific methylation epigenetic modification anti-tumor molecule and use thereof
  • pdl-1 gene target-specific methylation epigenetic modification anti-tumor molecule and use thereof
  • pdl-1 gene target-specific methylation epigenetic modification anti-tumor molecule and use thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0026] 1. Design and synthesis of human PD-L1 gene promoter-specific binding protein

[0027] Using ZF Tools Ver3.0 software, according to the CpG site of human PD-L1 promoter region, select the best sequence and design specific binding protein. The amino acid sequence of the selected DNA binding and mediating DNA methylation protein PPLA targeting the human PD-L1 promoter region is shown in SEQ.ID.No.1, and the nucleotide sequence of each corresponding DNA domain is SEQ. ID.No.2 shown.

[0028] 2. Construction of human PD-L1 gene promoter targeting methylase and adenovirus packaging

[0029] The artificially synthesized DNA fragment encoding PPLA was cloned and recombined into pcDNA3.1 (such as figure 1 ), and adenoviral packaging of the expression plasmid (such as figure 2 ), obtained the expressed genetically engineered protein PPLA, and confirmed its protein amount at 130KDa by Western-blot (such as image 3 ), through bioinformatics analysis to simulate the spatial ...

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Abstract

The invention belongs to the technical field of gene therapy, and specifically relates to an anti-tumor molecule for PDL-1 gene targeting-specific methylation epigenetic modification and application thereof. According to the principle of bioinformatics, epigenetic engineering and immunology, the present invention designs a new protein molecule PPLA which is modified by epimethylation for the promoter sequence of human PD‑L1 gene, and introduces it into human cells through an adenovirus vector. Highly express the protein molecule PPLA specific to the promoter sequence of the human PD‑L1 gene, and the expressed gene can effectively inhibit the PD‑L1 gene. This molecule can produce methylation chemical modification on the PDL‑1 gene. Internal experiments have confirmed that it has the function of efficiently blocking the expression of PDL‑1 gene, thereby breaking immune tolerance, and finally achieving the effect of treating chronic viral infection and tumor, and realizing the effect of anti-tumor and anti-viral immunotherapy. In the preparation of epigenetic engineering therapy It has broad application prospects in tumor and antiviral drugs.

Description

technical field [0001] The invention belongs to the technical field of gene therapy, and specifically relates to PDL-1 gene target-specific methylation epigenetic modification anti-tumor molecules and applications thereof. Background technique [0002] Immunity usually refers to the resistance of organisms to infectious diseases, the recognition of self and non-self substances, and the monitoring and removal of denatured substances produced in the body. Immunity is divided into innate immunity (innate immunity) and acquired immunity (acquired immunity), which can be subdivided into cellular immunity and humoral immunity. Immune regulation is an important part of immunity. Immunoregulation refers to the execution and rational regulation of the immune response process and level by the organism to meet the needs of immune defense and maintain the homeostasis of the organism. Its essence is a regulatory process involving multiple factors under the regulation of genetic genes. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C07K19/00C12N15/62C12N15/861A61K38/45A61K47/64A61P35/00A61P31/12
CPCA61K38/00C07K2319/00C07K2319/74C12N9/1007C12N15/86C12N2710/10343C12N2800/107
Inventor 朱乃硕李雪
Owner FUDAN UNIV