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Method for preparing direct composite PEI magnetic gene vector

A gene carrier and direct technology, applied in the field of carrier preparation, can solve the problems such as the preparation method of magnetic gene carrier that has not yet been seen, and achieve the effect of shortening the transfection time and improving the transfection efficiency

Inactive Publication Date: 2010-06-23
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Found through literature search to prior art, do not see the report relevant with the subject of the present invention " the preparation method of the magnetic gene carrier of direct composite PEI "

Method used

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  • Method for preparing direct composite PEI magnetic gene vector
  • Method for preparing direct composite PEI magnetic gene vector
  • Method for preparing direct composite PEI magnetic gene vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The materials involved in this embodiment are as follows:

[0027] Ferrous chloride, ferric chloride and sodium hydroxide are all analytically pure, purchased from China Pharmaceutical Group Shanghai Chemical Reagent Company;

[0028] African green monkey kidney cell line COS-7 cells were purchased from ATCC;

[0029] RPMI-1640 medium and serum were purchased from GIBICO;

[0030] The luciferase reporter plasmid pGL3-control was purchased from Promega, USA;

[0031] The PCS plasmid is disclosed in the publication specification of Chinese invention patent application CN101095951;

[0032] The luciferase detection kit was purchased from Promega, USA;

[0033] PEI-25K was purchased from Sigma;

[0034] BCA protein quantification kit was purchased from Shanghai Karma Biotechnology Company;

[0035] The preparation process of the magnetic gene carrier is as follows,

[0036] Take 5.6mmol FeCl 3 , 2.8mmol FeCl 2 Dissolve in 7ml of degassed water to get iron salt solut...

Embodiment 2

[0060] The materials involved in this example are the same as in Example 1, and the construction process of the magnetic gene carrier is also the same as in Example 1, except that the titration is stopped when the pH value is 10.5, and the stirring reaction in an ice bath is continued for 30 minutes;

Embodiment 3

[0062] The materials involved in this example are the same as in Example 1, and the construction process of the magnetic gene carrier is also the same as in Example 1, except that the titration is stopped when the pH value is 10.8, and the stirring reaction in an ice bath is continued for 35 minutes;

[0063]The particle size of the magnetic particles prepared in Example 2 and Example 3 is about 1um, and the particle itself is slightly red, indicating that the reaction is not complete, and the magnetic properties of the magnetic particles are slightly weak. After compounding with PEI25K, the resulting magnetic gene carrier MP- Compared with the magnetic gene carrier obtained in Example 1, PEI has no big difference in particle size and potential.

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Abstract

The invention relates to a method for preparing direct composite PEI magnetic gene vector in the biological technical field. The method comprises the steps: Step 1: FeCl3 and FeCl2 is taken to prepare magnetic particles with the coprecipitation method; Step 2: magnetic particles are compounded with polyethylene imine to obtain the magnetic gene vector. The magnetic gene vector prepared by the invention has stronger DNA bonding force and can carry exogenous genes to transfect exogenotes. Under the action of externally applied magnetic field, the transfection efficiency is increased obviously by 5 to 10 times and the transfection time is reduced to 1 hour.

Description

technical field [0001] The invention relates to a method for preparing a carrier in the field of biotechnology, in particular to a method for preparing a magnetic gene carrier directly compounded with PEI. Background technique [0002] As a new treatment for hereditary and infectious diseases, gene therapy has attracted more and more attention in recent years. However, various challenges and difficulties in the process of developing safe and effective gene carriers limit the clinical application of this treatment. Two-thirds of clinical trials use viral vectors, but they have many defects (C.E.Thomas, A.Ehrhardt, M.A.Kay, Progress and problems with the use of viral vectors for genetherapy. Nat.Rev.Genet.4, 2003, 346 ~358), such as high immunogenicity, potential tumorigenicity, inability to be used repeatedly in vivo, high price and so on. Therefore, people began to focus on the research and development of safe, low-toxic, and easy-to-operate non-viral gene vectors. Most n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85
Inventor 徐宇虹郭微
Owner SHANGHAI JIAO TONG UNIV