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Preparation method of carrier for gene transfer

A gene delivery and carrier technology, applied in the field of non-viral carrier-gelatin-siloxane nano-hybrid materials and its preparation, can solve the problems of non-viral nanoparticle gene carrier instability and low transfection rate, and achieve non-viral Immunogenicity, no side effects, good biocompatibility

Inactive Publication Date: 2011-03-30
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the instability and low transfection rate of non-viral nanoparticle gene carriers need to be solved before practical application.

Method used

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  • Preparation method of carrier for gene transfer
  • Preparation method of carrier for gene transfer
  • Preparation method of carrier for gene transfer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Dissolve 0.2g of gelatin in 20mL of acetic acid solution with a pH of 3, and stir at 50°C for 30min. Then 0.2 g of 3-(2,3-glycidoxy)propyltrimethoxysilane was added to the above 1% gelatin solution. After the reaction solution was stirred and reacted at 30° C. for 6 hours, ammonia water was added to adjust the pH value of the solution to 9, and the stirring reaction was continued for 12 hours to obtain a milky white suspension. The milky white suspension was subjected to high-speed centrifugation (20,000 rpm, 15 min, 30° C.) to obtain a white precipitate. The white precipitate was washed twice with distilled water to obtain figure 2 (a) Gelatin-siloxane nanohybrid material with a size of about 150 nm is shown. Characterized by infrared spectroscopy, such as figure 1 As shown, 1650cm -1 It is the stretching vibration absorption peak of carbonyl-C=O, 1541cm -1 is amino-NH 2 Bending vibrations, both derived from gelatin molecules, at 1030 and 1140 cm -1 is the str...

Embodiment 2

[0035] Dissolve 0.02g of gelatin in 20mL of hydrochloric acid solution with pH=3, and stir at 50°C for 30min. Then 0.2 g of 3-(2,3-glycidoxy)propyltrimethoxysilane was added to the above 0.1% gelatin solution. After the reaction solution was stirred and reacted at 50° C. for 30 minutes, 3-(trimethoxysilane) propylamine was added to adjust the pH value of the solution to 9, and the stirring reaction was continued for 12 hours to obtain a milky white suspension. The milky white suspension was subjected to high-speed centrifugation (20,000 rpm, 15 min, 30° C.) to obtain a white precipitate. The white precipitate was washed twice with distilled water to obtain figure 2 (b) shows a gelatin-siloxane nanohybrid material with a size of about 600 nm.

Embodiment 3

[0037]Dissolve 0.5g of gelatin in 20mL of hydrochloric acid solution with pH=3, and stir at 50°C for 30min. Then 0.25 g of 3-(2,3-glycidoxy)propyltrimethoxysilane was added to the above 2.5% gelatin solution. After the reaction solution was stirred and reacted at 50° C. for 30 minutes, 3-(trimethoxysilane) propylamine was added to adjust the pH to 10.0, and the stirring reaction was continued for 12 hours to obtain a milky white suspension. The milky white suspension was subjected to high-speed centrifugation (20,000 rpm, 15 min, 30° C.) to obtain a white precipitate. The white precipitate was washed twice with distilled water to obtain figure 2 (c) Gelatin-siloxane nanohybrid material with a size of 20 nm is shown.

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Abstract

The invention relates to a carrier for gene delivery, gelatin-siloxane hybrid nano-material, a preparation method thereof by two-step sol-gel reaction and an application thereof. The gelatin-siloxane nano-particles are prepared from gelatin and 3-(2,3-epoxy propoxy )propyltrimethoxysilicane by performing chemical combination and hydrolytic reaction in acidic condition to obtain sol; and adjustingthe pH value to slightly alkaline for performing polymerization reaction of silicane. The gelatin-siloxane hybrid nano-material has good biocompatibility, no toxic and side effect, and high stability; and is combined with exogenous gene to realize DNA transportation and expression, with high transduction efficiency, no immunogenicity, and no cytotoxicity. The size of the transferable exogenous gene is of several tens bp to several thousands kb, which overcomes size restriction to viral vector inserting into the exogenous gene. Thus the exogenous gene can be effectively carried into a host cell for performing effective transfection and expression.

Description

technical field [0001] The invention relates to a gene delivery carrier, in particular to a non-viral carrier-gelatin-siloxane nanometer hybrid material and its preparation method and application. Background technique [0002] Gene therapy refers to the biomedical technology that introduces human normal genes or genes with therapeutic effects into human target cells in a certain way to correct gene defects or play a therapeutic role, so as to achieve the purpose of treating diseases. Gene therapy has promising applications in replacing dysfunctional genes and in tumor therapy. Gene therapy has three basic elements: target gene, expression vector and delivery vector. The construction and improvement of delivery vector has always been the focus of gene therapy research. There are two types of delivery vectors: viral vectors and non-viral vectors. The transfection efficiency of viral vectors is high, but there are also some obvious disadvantages, such as high immunogenicity, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/42A61K48/00C12N15/63
Inventor 任磊赵阳王祖勇张其清
Owner XIAMEN UNIV
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