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Carrier for gene transmission and preparation method thereof and application

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: 2008-10-15
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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  • Carrier for gene transmission and preparation method thereof and application
  • Carrier for gene transmission and preparation method thereof and application
  • Carrier for gene transmission and preparation method thereof and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 0.2 g of gelatin was dissolved in 20 mL of acetic acid solution with pH 3 and stirred at 50° C. for 30 min. Then 0.2 g of 3-(2,3-glycidoxy)propyltrimethoxysilane was added to the above 1% gelatin solution. After the reaction solution was stirred at 30° C. for 6 hours, ammonia water was added to adjust the pH 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 the following figure 2 (a) Gelatin-siloxane nanohybrid material with a size of approximately 150 nm shown. Characterized by infrared spectrum, such as figure 1 shown, 1650cm -1 is the stretching vibration absorption peak of carbonyl-C=O, 1541cm -1 is amino-NH 2 Bending vibrations, both derived from gelatin molecules, 1030 and 1140 cm -1 is the stretchin...

Embodiment 2

[0035] 0.02 g of gelatin was dissolved in 20 mL of hydrochloric acid solution with pH=3, and stirred at 50° C. for 30 min. 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 at 50° C. for 30 min, 3-(trimethoxysilane)propylamine was added to adjust the pH of the solution to 9, and the reaction was continued to be stirred 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 the following figure 2 (b) Gelatin-siloxane nanohybrid material with a size of about 600 nm shown.

Embodiment 3

[0037]0.5 g of gelatin was dissolved in 20 mL of hydrochloric acid solution with pH=3, and stirred at 50° C. for 30 min. 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 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 the following figure 2 (c) Gelatin-siloxane nanohybrid material with a size of 20 nm 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 adjusting the 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 present invention relates to a gene delivery carrier, in particular to a non-viral carrier-gelatin-siloxane nano-hybrid material and a preparation method and application thereof. Background technique [0002] Gene therapy refers to a biomedical technology that introduces normal human 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 shortcomings, such as high immunogeni...

Claims

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Application Information

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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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