Cationized polysaccharide nanoparticle gene delivery systems and manufacturing method thereof

A gene delivery system and cationization technology, applied in the field of cationized polysaccharide nanoparticle gene delivery system, can solve the problems of no positive charge and cannot be directly used as a gene, etc., and achieve good encapsulation and release effect, no immunogenicity, strong effect of proliferation

Inactive Publication Date: 2012-11-07
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, except for chitosan, other natural plant polysaccharides are almost not positively charged, so they cannot be directly used as gene carriers. Natural polysaccharides must be cationized and modified to increase their potential before they can successfully bind to DNA plasmids.

Method used

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  • Cationized polysaccharide nanoparticle gene delivery systems and manufacturing method thereof
  • Cationized polysaccharide nanoparticle gene delivery systems and manufacturing method thereof
  • Cationized polysaccharide nanoparticle gene delivery systems and manufacturing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Preparation of the gene delivery system of the cationized Pleurotus eryngii polysaccharide-DNA plasmid nanocomposite modified by spermine

[0056] Weigh 0.5g refined Pleurotus eryngii polysaccharide, dissolve in 30ml double distilled water, add 0.75g KIO 4 , quickly placed in a dark room, stirred magnetically, and reacted at room temperature for 72 hours; adding 10ml ethylene glycol to the reaction solution to terminate the reaction, and continued the reaction for 30 minutes according to the above conditions; put the reaction solution into a dialysis bag (cutoff molecular weight > 3500Da), and dialyzed in double distilled water for 48 hours ; The dialysate was freeze-dried to obtain the oxidized Pleurotus eryngii polysaccharide.

[0057] Take 0.15g of the above-mentioned oxidized Pleurotus eryngii polysaccharide and dissolve it in 10ml of double distilled water; weigh 0.3g of spermine and dissolve it in 5ml of borate buffer solution (pH=9); use a disposable...

Embodiment 2

[0059] Example 2: Preparation of a gene delivery system of ethylenediamine-modified cationized Pleurotus eryngii polysaccharide-DNA plasmid nanocomposite

[0060] Weigh 0.6 g refined Pleurotus eryngii polysaccharide, dissolve in 50ml double distilled water, add 0.89g KIO 4 , quickly placed in a dark room, magnetically stirred, and reacted at room temperature for 72 hours; adding 12ml of ethylene glycol to the reaction solution to terminate the reaction, and continued the reaction for 30 minutes according to the above conditions; put the reaction solution into a dialysis bag and dialyzed in double distilled water for 48 hours (molecular weight cut-off > 3500Da) ; The dialysate was freeze-dried to obtain the oxidized Pleurotus eryngii polysaccharide.

[0061] Take 0.4 g of oxidized Pleurotus eryngii polysaccharide and dissolve it in 30ml of double distilled water; take 0.7 ml of ethylenediamine and dissolve it in 5ml of borate buffer solution (pH=9); use a disposable syringe t...

Embodiment 3

[0063] Example 3: Preparation of the gene delivery system of cationized mulberry leaf polysaccharide-DNA plasmid nanocomposite modified by ethylenediamine

[0064] Weigh 0.8g refined mulberry leaf polysaccharide, dissolve in 50ml double distilled water, add 1.2g KIO 4 , quickly placed in a dark room, magnetically stirred, and reacted at room temperature for 72 hours; adding 12ml of ethylene glycol to the reaction solution to terminate the reaction, and continued the reaction for 30 minutes according to the above conditions; put the reaction solution into a dialysis bag and dialyzed in double distilled water for 48 hours (molecular weight cut-off > 3500Da) ; The dialysate was freeze-dried to obtain the oxidized mulberry leaf polysaccharide.

[0065] Take 0.5g of oxidized mulberry leaf polysaccharide and dissolve it in 30ml of double distilled water; take 0.28ml of ethylenediamine and dissolve it in 5ml of borate buffer solution (pH=9); Add it into the mulberry leaf polysacch...

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Abstract

The invention discloses cationized polysaccharide nanoparticle gene delivery systems, which are gene delivery systems combining polysaccharides modified by an amine compound and DNA plasmids, wherein the mass ratio of the cationized polysaccharides to the DNA plasmids is (0.5-200):1; the particle size of the cationized polysaccharide-DNA plasmid nano composites is 21 to 414 nanometers; and the amine compound is spermine, ethylenediamine or polyethyleneimine with a number-average molecular weight of 600Da to 2,000Da. The polysaccharides of traditional Chinese medicine universally have various bioactivities for immunoregulation, aging resistance, coagulation resistance and the like. Compared with other viral vector and other non-viral vector gene delivery systems, the cationized polysaccharide nanoparticle gene delivery systems are safe, free from immunogenicity and biodegradable, and the preparation process of the cationized polysaccharide nanoparticle gene delivery systems is simple and economic. The cationized polysaccharides have good DNA plasmid bonding actions and gene delivery and expression functions. The positive charges on the aminos bonded with the polysaccharide chains can effectively combine with the negative charges on the DNA plasmids to protect the plasmids from being degraded by various enzymes in and out cells.

Description

technical field [0001] The invention relates to the field of plant functional polysaccharides and a gene delivery system, in particular to a cationic polysaccharide nanoparticle gene delivery system. Background technique [0002] Gene therapy has opened up its broad application prospects in the field of biomedicine, and can be used to treat genetic and acquired diseases such as hemophilia, cystic fibrosis, gynecological diseases, etc. [See: Jay Lozier. Gene therapy of the hemophilias. Seminars in Hematology, 2004, 41 (4): 287~296. Uta Griesenbach, A. Christopher Boyd. Pre-clinical and clinical endpoint assays for cystic fibrosis gene therapy. Journal of Cystic Fibrosis, 2005, 4 (2): 89~ 100. Memy H, Hassan, Essam E, Othman, Daniela Hornung, Ayman A1-Hendy. Gene therapy of benign gynecological diseases. Advanced Drug Delivery Reviews, 2009, 61(10):822~835.]. The key to gene therapy technology is to be able to effectively deliver foreign genes into the nucleus and make them e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63B82Y5/00
Inventor 徐希明曹霞余江南王淼邓纹纹
Owner JIANGSU UNIV
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