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Construction and applications of multifunctional graphene gene vector

A gene carrier and graphene technology, applied in other methods of inserting foreign genetic materials, recombinant DNA technology, etc., can solve the problems of cell damage, cell hemolysis, cytotoxicity, etc., and achieve reduced toxicity, high transfection efficiency, and toxicity low effect

Active Publication Date: 2018-03-16
NANJING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, PEI itself has a large number of cationic groups, which will cause a certain degree of damage to the cell membrane and cause hemolysis of the cells, so it has certain cytotoxicity and will inevitably damage the cells during the transfection process.

Method used

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  • Construction and applications of multifunctional graphene gene vector
  • Construction and applications of multifunctional graphene gene vector
  • Construction and applications of multifunctional graphene gene vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Functionalized graphene quantum dots are used for gene transfection

[0040] To prepare functionalized graphene quantum dots, other implementation steps were the same as in Comparative Example 1 except that branched PEI was added to change it to a branched PEI with a molecular weight of 25,000. Transmission scanning electron microscopy images of functionalized graphene quantum dots such as figure 1 , it can be seen that the functionalized graphene quantum dots are more uniformly dispersed. And the fluorescence test images of functionalized graphene quantum dots are as follows figure 2 , proving that functionalized graphene quantum dots have good fluorescence properties. The optimal transfection efficiency was determined to be 51.63% by flow cytometry, and the transfection images were observed with a confocal microscope as follows: image 3A-1, the dotted part of the hash distribution is the part of the transfected cells, and the transfected part can be se...

Embodiment 2

[0041] Example 2: Functionalized graphene quantum dots are used for gene transfection

[0042] To prepare functionalized graphene quantum dots, the implementation steps are the same as in Example 1 except that the amount of branched PEI added is increased to 3 mg. The optimal transfection efficiency was determined to be 80.57% by flow cytometry, and the transfection image was observed with a confocal microscope as follows: image 3 In B-1, the dotted part of the hash distribution is the part of the transfected cells, and the transfected part can be seen to be about 80% of the cells by naked eyes. The bright-field image is 3B-2, and the cos7 cells can be seen to have a clear outline and a relatively uniform distribution of cells. Cytotoxicity test results such as Figure 4 In Example 2, the cell survival rate was 89.34%.

Embodiment 3

[0043] Example 3: Functionalized graphene quantum dots are used for gene transfection

[0044] To prepare functionalized graphene quantum dots, the implementation steps are the same as in Example 1 except that the amount of branched PEI added is increased to 5 mg. The optimal transfection efficiency was determined to be 74.13% by flow cytometry, and the transfection images were observed with a confocal microscope as follows: image 3 In C-1, the dotted part of the hash distribution is the part of the transfected cells, and the transfected part can be seen to be about 70% of the cells by naked eyes. The bright-field image is 3C-2, and the outline of cos7 cells can be seen clearly, and the cells are evenly distributed. Cytotoxicity test results such as Figure 4 In Example 3, the cell survival rate was 87.85%.

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Abstract

The invention discloses a multifunctional graphene gene vector construction method, wherein small-size graphene quantum dot with fluorescence is used as a basic vector, and is functionally linked to acationic polymer branched chain polyethyleneimine so as to achieve gene transfection ability, and the non-gene-transfection cationic polymer part can be tracked due to the fluoresce ability. According to the present invention, the synthesis process is simple, the constructed multifunctional graphene gene vector has advantages of low toxicity and high transfection efficiency, the highest transfection efficiency can achieve 80.57%, gene transfection and tracking can be simultaneously achieved, and the constructed multifunctional graphene gene vector can be used for cell biology research and gene therapy field.

Description

technical field [0001] The invention belongs to the field of inorganic chemistry and biomedicine, and relates to the construction and application of a multifunctional graphene gene carrier, in particular to a simple, efficient, and low-toxicity modified polyethylenimine applied to gene transfection and tracer Multifunctional graphene gene carrier. Background technique [0002] As nano-sized graphene, graphene quantum dots not only have the excellent properties of graphene itself, but also exhibit unique photochemical properties due to the edge effect and small size effect, and exhibit low cytotoxicity, excellent solubility and good cell compatibility. In addition, the edges of graphene quantum dots usually contain carboxyl and hydroxyl groups, which can interact with biomolecules. [0003] Gene transfection technology is a technology that delivers purified exogenous DNA and RNA into cells and expresses them in cells. Gene transfection vectors are divided into two categori...

Claims

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Application Information

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IPC IPC(8): C12N15/87
CPCC12N15/87
Inventor 董伟盛伟陈琦李军舰左淦丞
Owner NANJING UNIV OF SCI & TECH
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