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Artificial zinc finger protein transcription factor capable of starting A20 gene expression and use

A zinc finger protein and transcription factor technology, applied in genetic engineering, plant genetic improvement, animal/human peptides, etc., can solve the problems of artificial zinc finger protein transcription factors, etc., to suppress organ transplant rejection, avoid side effects, The effect of wide application prospects

Inactive Publication Date: 2008-12-31
ARMY MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are currently no research reports on the construction of artificial zinc finger protein transcription factors using the A20 gene as a drug target at home and abroad.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1. An artificial zinc finger transcription factor protein that can initiate the expression of A20 gene in vivo. Its protein structure includes a binding domain, a functional domain, a nuclear localization signal and a transmembrane domain; its binding domain is the target sequence of the promoter region of the A20 gene ( 18bp) specific binding zinc finger motif polypeptide, wherein: the target sequence (18bp) of the promoter region of the binding domain A20 gene is SEQ ID NO:1;

[0051] The zinc finger motif polypeptide ZFP1 that specifically binds is the following three short peptides, which are connected by TGEKP to form a fusion polypeptide. The N-terminal amino acid sequence of the fusion polypeptide is LEPGEKP-, and the C-terminal is TGKKTS: the three short peptides The sequence of the zinc finger motifs of the peptides is:

[0052] SEQ ID NO: 2 QSGDLRR, SEQ ID NO: 3 SKKHLAE, SEQ ID NO: 4Q SGNLTE

[0053] The above-mentioned zinc finger motif N-terminal sequenc...

Embodiment 2

[0055] Example 2. An artificial zinc finger transcription factor protein that can initiate the expression of A20 gene in vivo. Its protein structure includes a binding domain, a functional domain, a nuclear localization signal and a transmembrane domain; its binding domain is the target sequence of the promoter region of the A20 gene ( 18bp) specific binding zinc finger motif polypeptide, wherein: the target sequence (18bp) of the promoter region of the binding domain A20 gene is SEQ ID NO:1;

[0056] The zinc finger motif polypeptide ZFP2 that specifically binds is the following four short peptides, which are connected by TGEKP to form a fusion polypeptide. The N-terminal amino acid sequence of the fusion polypeptide is LEPGEKP-, and the C-terminal is TGKKTS: the four short peptides The sequence of the zinc finger motifs of the peptides is:

[0057] SEQ ID NO: 2 QSGDLRR, SEQ ID NO: 3 SKKHLAE, SEQ ID NO: 4 QSGNLTE

[0058] SEQ ID NO: 5 QNSTLTE

[0059] The above-mentioned zinc fin...

Embodiment 3

[0061] Example 3. An artificial zinc finger transcription factor protein that can initiate the expression of A20 gene in vivo. Its protein structure includes a binding domain, a functional domain, a nuclear localization signal, and a transmembrane domain; its binding domain is the target sequence of the promoter region of the A20 gene ( 18bp) specific binding zinc finger motif polypeptide, wherein: the target sequence (18bp) of the promoter region of the binding domain A20 gene is SEQ ID NO:1;

[0062] The zinc finger motif polypeptide ZFP3 that specifically binds is the following five short peptides, which are connected by TGEKP to form a fusion polypeptide. The N-terminal amino acid sequence of the fusion polypeptide is LEPGEKP-, and the C-terminal is TGKKTS: the five short peptides The sequence of the zinc finger motifs of the peptides is:

[0063] SEQ ID NO: 2 QSGDLRR, SEQ ID NO: 3 SKKHLAE, SEQ ID NO: 4 QSGNLTE

[0064] SEQ ID NO: 5 QNSTLTE, SEQ ID NO: 6 DPGHLVR

[0065] The a...

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PUM

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Abstract

The invention relates to an artificial zinc finger protein transcription factor which can activate the A20 gene expression, and the application of the artificial zinc finger protein transcription factor. The preparation method comprises the preparation of a zinc finger motif polypeptide bonded with the A20 gene promoter region target sequence specificity, as well as the synthesis of an artificial transcription factor; the artificial transcription factor comprises a binding region, a functional region, a nuclear localization signal and a transmembrane region; the connection order is the transmembrane region-the nuclear localization signal - the binding region - the functional region; the prepared polypeptide containing 3 to 6 zinc finger sequences is syncretized into the functional region of NF-p65 or VP64 and joined with the nuclear localization signal; 9 arginines or 12 amino acids of a Tat protein are added to the N-terminal to serve as a protein transduction region. The invention can effectively span the cell membranes to enter the cells to activate the expression of the A20 gene, and has good anti-vascular restenosis, atherosclerosis sclerosis, cardiac hypertrophy, serious inflammatory injury and other performances; the invention can be applied to the preparation of the cell protection drugs for overcoming the rejection of organ transplants or cardiac hypertrophy or serious inflammatory injury.

Description

Technical field [0001] The invention relates to an artificial zinc finger protein transcription factor capable of initiating A20 gene expression and its use. technical background [0002] The A20 gene belongs to the zinc finger protein family. Gene knockout experiments show that the A20 gene is an important functional gene that is indispensable for the body to resist inflammation. Expression of A20 gene in islet cells can effectively induce islet transplantation tolerance, and overexpression of A20 gene can prevent the formation of graft atherosclerosis in all kinds of kidney transplantation. The A20 gene can inhibit the CD40-CD40 ligand signaling pathway, thereby inhibiting the formation of atherosclerosis and the occurrence of graft atherosclerosis. The A20 gene can dynamically manage the heart and inhibit myocardial hypertrophy. The A20 gene can effectively inhibit the intimal hyperplasia caused by the pathological proliferation and migration of smooth muscle cells, resist end...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12A61K38/17A61P37/06A61P9/10
Inventor 朱楚洪应大君魏勇侯春丽
Owner ARMY MEDICAL UNIV
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