Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for real-time monitoring on gene transfection

A real-time monitoring and gene transfection technology, applied in the field of genetic engineering, can solve the problems of high cost and complicated process, and achieve the effect of low cost, mild conditions, and easy operation and promotion

Inactive Publication Date: 2013-05-29
SHENZHEN INST OF ADVANCED TECH
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the traditional methods for visually tracing the distribution and localization of exogenous transfected genes in cells mainly focus on the application of nanomaterials with optical characteristics for labeling, the process is complicated and the cost is relatively high

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
  • Method for real-time monitoring on gene transfection
  • Method for real-time monitoring on gene transfection
  • Method for real-time monitoring on gene transfection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Synthesis of tetramethylrhodamine isothiocyanate-labeled polyethyleneimine:

[0036] ① Configure a dimethyl sulfoxide solution of polyethyleneimine with a concentration of 17.86mM;

[0037] ②A solution of tetramethylrhodamine tetramethylisothiocyanate in dimethyl sulfoxide with a concentration of 10 mM is prepared;

[0038] ③ According to the molar ratio of polyethyleneimine and tetramethylrhodamine tetramethylisothiocyanate, the dimethyl sulfoxide solution of polyethyleneimine and tetramethylrhodamine tetramethylisothiocyanate were mixed in a ratio of 1:10. The sulfoxide solution was mixed and stirred for 6 hours. After 6 hours, the mixed solution was transferred to a dialysis bag with a molecular weight cut-off of 3500. First, it was dialyzed three times with dimethyl sulfoxide for one day, and then continued to dialyze with ultrapure water for two days. sky.

Embodiment 2

[0039] Embodiment 2: the synthesis of the polyethylenimine of tetramethyl rhodamine isothiocyanate label:

[0040] ① Configure a dimethyl sulfoxide solution of polyethyleneimine with a concentration of 17.86mM;

[0041] ②A solution of tetramethylrhodamine tetramethylisothiocyanate in dimethyl sulfoxide with a concentration of 10 mM is prepared;

[0042] ③ According to the molar ratio of polyethyleneimine and tetramethylrhodamine tetramethylisothiocyanate, the dimethyl sulfoxide solution of polyethyleneimine and tetramethylrhodamine tetramethylisothiocyanate were mixed in a ratio of 1:20 The sulfoxide solution was mixed and stirred for 6 hours. After 6 hours, the mixed solution was transferred to a dialysis bag with a molecular weight cut-off of 3500. It was dialyzed 5 times with dimethyl sulfoxide for 2 days, and then continued with ultrapure water for 3 days. sky.

Embodiment 3

[0043] Embodiment 3: the synthesis of the polyethylenimine of tetramethylrhodamine isothiocyanate label:

[0044]Embodiment 2: the synthesis of the polyethylenimine of tetramethyl rhodamine isothiocyanate label:

[0045] ① Configure a dimethyl sulfoxide solution of polyethyleneimine with a concentration of 17.86mM;

[0046] ②A solution of tetramethylrhodamine tetramethylisothiocyanate in dimethyl sulfoxide with a concentration of 10 mM is prepared;

[0047] ③ According to the molar ratio of polyethyleneimine and tetramethylrhodamine tetramethylisothiocyanate, the dimethyl sulfoxide solution of polyethyleneimine and tetramethylrhodamine tetramethylisothiocyanate were mixed in a ratio of 1:40. The sulfoxide solution was mixed and stirred for 6 hours. After 6 hours, the mixed solution was transferred to a dialysis bag with a molecular weight cut-off of 3500. First, it was dialyzed 4 times with dimethyl sulfoxide for 1.5 days, and then continued to dialyze with ultrapure water fo...

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

No PUM Login to View More

Abstract

The invention relates to a method for real-time monitoring on gene transfection. Based on polyethyleneimine marked by tetramethyl isothiocyanic acid rhodamine, polyethyleneimine is subjected to tetramethyl isothiocyanic acid rhodamine marking at a room temperature. The method has the advantages of simplicity of synthesis, moderate conditions, low cost and convenience for operation. Plasmid DNA (Deoxyribonucleic Acid) of a fluorescent protein is expressed by electrostatic adsorption of the polyethyleneimine marked by the tetramethyl isothiocyanic acid rhodamine, and the plasmid DNA is transfected into cells; a fluorescence imaging method is used for monitoring the distribution of the polyethyleneimine which is used as a carrier and a plasmid in the cells and the expression of the plasmid in the cells in real time; and the method is simple and easy to realize, is suitable for most of laboratories and is convenient for operation and popularization.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for real-time monitoring of gene transfection. Background technique [0002] With the completion of the Human Genome Project and the advancement of cell biology technology, gene therapy will occupy an important position in the process of overcoming the stubborn disease of human beings, and will become a common treatment method. Gene therapy refers to a new 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 exert therapeutic effects, so as to achieve the purpose of treating diseases. Gene transfection technology has been widely used in gene therapy research. Gene transfection refers to the transfer or transport of nucleic acid molecules (genes) with biological functions into cells and the maintenance of biological functions of nucleic acid molecules (genes) in ce...

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): C12Q1/68C12Q1/02C08G73/04
Inventor 蔡林涛石碧华龚萍刘朋王碧王亚楠
Owner SHENZHEN INST OF ADVANCED TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com