Unlock instant, AI-driven research and patent intelligence for your innovation.

Gene transporter as well as preparation method and application thereof

A transporter and gene technology, applied in the field of medicine, can solve problems such as inappropriate embryo cells, and achieve the effects of low toxicity, high stability and small particle size

Active Publication Date: 2020-05-15
FUYANG NORMAL UNIVERSITY
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, based on the specificity of embryonic cells, the vectors provided by these traditional techniques are not suitable for embryonic cells

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Gene transporter as well as preparation method and application thereof
  • Gene transporter as well as preparation method and application thereof
  • Gene transporter as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0036] The preparation method of the above-mentioned gene transporter, the preparation method comprises: adding the nano-metal particle solution to the solution of the polyethyleneimine, stirring, and the self-assembly of the polyethyleneimine and the nano-metal particle to form a The gene transporter of the nucleocapsid structure.

[0037] Preferably, the mass percent content of polyethyleneimine in the polyethyleneimine solution is 0.01-1%, and the mass percent content of nano-metal particles in the nano-metal particle solution is 1-3%.

[0038] Preferably, the stirring temperature is 22-27°C, the stirring speed is 700-900r / min, and the stirring time is 45-50h.

[0039] Preferably, the preparation method is further subjected to membrane filtration after stirring, including but not limited to using a 220nm filter membrane. The obtained gene transporter can be stored at low temperature, including but not limited to 4°C.

[0040] The embodiment of the present invention also p...

Embodiment 1

[0050] 1 Materials, equipment and methods

[0051] 1.1 Materials

[0052] Polyethyleneimine (PEI, Sigma-Aldrich company, relative molecular mass: 25000, 1800), chloroauric acid (HAuCl 4 ·3H 2 O, Sigma-Aldrich Company, relative molecular mass 394), green fluorescent protein reporter gene plasmid EGFP (self-constructed), TUNEL cell apoptosis detection kit (Shanghai Beyond Biotechnology Co., Ltd.), Anti-phospho-H2A.X ( Merck Chemical Technology (Shanghai) Co., Ltd.), the rest of the reagents were of domestic analytical grade, and the water used in all experiments was ultrapure water.

[0053] 1.2 Equipment

[0054] Intelligent CNC temperature-controlled magnetic heating stirrer (model ZNCL-BS, Chongqing Dongyue Instrument Co., Ltd.), ultraviolet spectrophotometer (Thermo, the United States), agarose gel electrophoresis instrument (Bio-Rad Company, the United States), laser particle size analysis Instrument (Nano90, Malwern Company, UK), transmission electron microscope (JEOL-...

Embodiment 2

[0123] This example is a modification example of Example 1. The change relative to Example 1 is that in the step "1.3.1 Preparation of Gene Carrier AuNP-PEI", PEI with a molecular weight of 25,000 and PEI with a molecular weight of 1,800 were used and the two The mass ratio of the latter is 1:1.

[0124] See results Figure 12 ,according to Figure 12 , when the mass ratio of the first PEI and the second PEI is 1:1, the proportion of blastocysts showing fluorescent signals is also higher.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a gene transporter as well as a preparation method and application thereof. The gene transporter comprises nano metal particles and polyethyleneimine and the gene transporteris of a core-shell structure, wherein the nano metal particles form the core of the gene transporter; the polyethyleneimine comprises first polyethyleneimine of which the molecular weight is 20000-30000 and second polyethyleneimine of which the molecular weight is 1500-2000; and the first polyethyleneimine and the second polyethyleneimine wrap the surface of the core in an interweaving manner to form a wrapping shell. After being combined with a plasmid which is inserted into an exogenous target gene segment through an electrostatic adsorption function, the gene transporter is capable of breaking through a transparent tape barrier on the outer layer of an embryo cell plasma membrane and entering an embryo cell, and promoting breakage of double-chain DNA (deoxyribonucleic acid) of the embryo cell, so that conditions can be created for integration of the exogenous target gene segment. Meanwhile, the inventor finds that the gene transporter also has the advantageous effects of being smallin particle size, good in dispersibility, high in stability, low in toxicity, and the like.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a gene transporter and its preparation method and application. Background technique [0002] In experiments related to biological research, it is an important experimental method to introduce specific exogenous genes into mouse early embryo cells. There are various technical methods currently used, but there are some disadvantages, such as: the required instruments are precise and expensive, the technical threshold is high, physical and chemical damage to the embryo is caused, commercial transfection reagents are not applicable and expensive, etc. In addition, embryonic cells are different from somatic cells. There is a layer of zona pellucida on the outer layer of the cytoplasmic membrane. The double-layer biological barrier makes it difficult for foreign genes to enter embryonic cells. [0003] Metal nanomaterials have been widely used in gene therapy and biological research ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87
CPCC12N15/87
Inventor 张迪王旗李文雍王荣程小翠李欢欢
Owner FUYANG NORMAL UNIVERSITY