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Functional gene carrier and DNA/carrier compound

A gene carrier and functional technology, applied in the field of biomedical carrier materials, can solve problems affecting operability and reproducibility, lack of regulation in the process, and lower transfection efficiency to achieve excellent transfection ability, prolong cycle time, The effect of improving transfection efficiency

Inactive Publication Date: 2018-05-22
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, during the preparation, due to the lack of control in the process of generating the carrier after the mixture of calcium ions and carbonate ions, it is easy to cause the particle size of the carrier to be too large, thereby reducing the transfection efficiency
At the same time, it also affects the operability and reproducibility of the carrier preparation process

Method used

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  • Functional gene carrier and DNA/carrier compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A functional gene carrier, composed of calcium carbonate, sodium alginate and polypeptide, the mass ratio of calcium carbonate, sodium alginate and polypeptide is 2:1:3, the molecular weight of the sodium alginate is 100kDa, the The polypeptide is PKKKRKVH7R8, and its amino acid sequence is shown in SEQ ID NO:1.

[0025] A DNA / carrier complex, consisting of a functional gene carrier and plasmid DNA (luciferase plasmid pGL-3), the mass ratio of the functional gene carrier to the plasmid DNA is 30:1.

[0026] The preparation method of the DNA / carrier complex is as follows:

[0027] (1) Prepare 1 mL of mixed aqueous solution of 100 mg / mL calcium chloride and 100 μg / mL plasmid DNA (luciferase plasmid pGL-3);

[0028] (2) Prepare 1 mL of a mixed aqueous solution of 1500 μg / mL polypeptide, 500 μg / mL sodium alginate and 1.06 mg / mL sodium carbonate;

[0029] (3) Add the solution of step (1) dropwise to the solution of step (2), so that calcium chloride and sodium carbonate re...

Embodiment 2

[0032] A functional gene carrier, composed of calcium carbonate, sodium alginate and polypeptide, the mass ratio of calcium carbonate, sodium alginate and polypeptide is 4:2:5, the molecular weight of the sodium alginate is 50kDa, the The polypeptide is PKKKRKVH7R8.

[0033] A DNA / carrier complex, consisting of a functional gene carrier and plasmid DNA (luciferase plasmid pGL-3), the mass ratio of the functional gene carrier to the plasmid DNA is 110:1.

[0034] The preparation method of the DNA / carrier complex is as follows:

[0035] (1) Prepare 1 mL of mixed aqueous solution of 200 mg / mL calcium chloride and 50 μg / mL plasmid DNA;

[0036] (2) Prepare 1 mL of a mixed aqueous solution of 2500 μg / mL polypeptide, 1000 μg / mL sodium alginate and 2.12 mg / mL sodium carbonate;

[0037] (3) Add the solution of step (1) dropwise to the solution of step (2), so that calcium chloride and sodium carbonate react to form a suspension solution of DNA / carrier complex.

[0038] The measurem...

Embodiment 3

[0040] A functional gene carrier, composed of calcium carbonate, sodium alginate and polypeptide, the mass ratio of calcium carbonate, sodium alginate and polypeptide is 1:1:1, the molecular weight of the sodium alginate is 150kDa, the The polypeptide is PKKKRKVH7R8.

[0041] A DNA / carrier complex, consisting of a functional gene carrier and plasmid DNA (luciferase plasmid pGL-3), the mass ratio of the functional gene carrier to the plasmid DNA is 2:1.

[0042] The preparation method of the DNA / carrier complex is as follows:

[0043] (1) Prepare 1 mL of mixed aqueous solution of 20 mg / mL calcium chloride and 300 μg / mL plasmid DNA;

[0044] (2) Prepare 1 mL of a mixed aqueous solution of 200 μg / mL polypeptide, 200 μg / mL sodium alginate and 0.212 mg / mL sodium carbonate;

[0045] (3) Add the solution of step (1) dropwise to the solution of step (2), so that calcium chloride and sodium carbonate react to form a suspension solution of DNA / carrier complex.

[0046] The measuremen...

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Abstract

The invention discloses a functional gene carrier, which is composed of calcium carbonate, sodium alginate, and polypeptide according to a mass ratio of 1-8:1-4:1-12. The invention also discloses a DNA / carrier compound, which has the advantages of low toxicity, high efficiency, stability, simple and convenient preparation process, and mild preparation conditions. The compound has a wide application prospect in the biomedicine field such as a gene / drug delivery system, and the like.

Description

technical field [0001] The invention relates to the field of biomedical carrier materials, in particular to a functional gene carrier, and also to a DNA / carrier complex and a preparation method thereof. Background technique [0002] With the acceleration of human exploration in the field of biomedicine, new breakthrough methods have been gradually discovered for some intractable diseases that have been treated with little effect by traditional chemical drugs, and gene therapy is one of the important theories Innovation and technological innovation. In the process of gene therapy, functional genes are delivered to target cells or tissues through various carriers to repair damaged / missing genes, so as to achieve the purpose of curing diseases. Among them, the construction and screening of suitable vectors to achieve effective gene delivery is crucial to the success of gene therapy. Vectors for gene delivery are divided into two categories: viral vectors and non-viral vectors...

Claims

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Application Information

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IPC IPC(8): C12N15/87
CPCC12N15/87
Inventor 瞿玮张先正曾旋谢书宇潘源虎
Owner HUAZHONG AGRI UNIV
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