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Histone simulated gene vector and preparation method and application thereof

A histone gene and carrier technology, applied in the field of gene carrier, can solve the problems of high price and difficulty in histone purification and separation, and achieve the effects of good biocompatibility, high-efficiency compression, and excellent transfection ability

Inactive Publication Date: 2015-11-18
CHINA PHARM UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the purification and separation of natural histones is difficult and expensive

Method used

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  • Histone simulated gene vector and preparation method and application thereof
  • Histone simulated gene vector and preparation method and application thereof
  • Histone simulated gene vector and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the preparation of PCAH gene carrier

[0030] Weigh 0.2 g of agmatine (1 mmol), 0.52 g of diacrylcystamide (2 mmol) and 0.18 g of histamine (1 mmol) in a flask, add 3 mL of methanol and stir and blend. Protected by Ar, the temperature of the oil bath was raised to 90°C, the reaction was stirred for 24 hours, and purified by dialysis. The NMR spectrum of the histone-like gene carrier PCAH is as follows Figure 1A As shown, its molecular weight is characterized by GPC as Figure 1B shown.

Embodiment 2

[0031] Embodiment 2: the preparation of the gene carrier system (PCAH-DNA) that PCAH gene carrier combines with DNA

[0032] The PCAH gene carrier and DNA were mixed in different mass ratios and incubated for 30 minutes to form the PCAH-DNA complex. The optimal binding mass ratio of the PCAH-DNA complex obtained from the gel electrophoresis experiment is 2:1, and the experimental results are as follows figure 2 . The particle diameter and surface charge of gained PCAH-DNA complex are as follows:

[0033] PCAH: DNA (w / w)

Embodiment 3

[0034] Embodiment 3: Reduction response experiment of PCAH-DNA gene carrier system

[0035] Dithiothreitol (DTT) was used to simulate the high-concentration GSH environment in the cell, and the reduction response ability of the PCAH-DNA gene carrier system was investigated by gel electrophoresis experiment. The ratio of each component in the experiment was as follows:

[0036] Polymer / DNA

[0037] The experimental results obtained are as image 3 shown.

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Abstract

The present invention discloses a histone simulated gene vector and a preparation method and application thereof, and in particular relates to a histone imitated gene vector PCAH capable of gene delivery. The invention also provides a gene vector system which is obtained through electrostatic adsorption self-assembly between nucleic acids and a polymer synthesized from arginine and histidine basic amino acid decarboxylated products, agmatine, histamine and bis(acryloyl)cystamine. The invention also discloses the preparation method of the histone simulated gene vector and application of the histone simulated gene vector to gene therapy. The histone simulated gene vector has good cell penetration function, high cell transfection efficiency and low cytotoxicity, and provides an effective mean for gene therapy of diseases.

Description

technical field [0001] The invention relates to a histone-like gene carrier and its preparation method and application, belonging to the technical field of gene carriers. Background technique [0002] Gene therapy provides a new possibility for the treatment of difficult diseases such as cancer. Successful gene therapy can successfully deliver the target gene to the diseased cells and enable the expression of the target gene. This process requires the help of a certain gene delivery carrier. Gene delivery vectors include viral vectors and non-viral vectors. Although the gene transfection efficiency of viral vectors is high, non-viral vectors have the characteristics of low immunogenicity, targeting specificity, low cost, and convenient use, which are superior to viral vectors in many aspects. In addition to the advantages of the biological carrier gene delivery system, the histone-like gene carrier system studied in this topic also has a significant increase in transfectio...

Claims

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Application Information

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IPC IPC(8): C12N15/87C08G69/42A61K47/34A61K48/00A61P35/00A61P3/00A61P25/00
Inventor 姜虎林崔朋飞邢磊庄婉茹
Owner CHINA PHARM UNIV
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