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Method for transferring exogenous gene into animal cell

A technology of exogenous genes and animal cells, applied to other methods of inserting foreign genetic materials, using microinjection methods, using vectors to introduce foreign genetic materials, etc., can solve problems such as difficulty in mastering, instability, and low birth rate

Inactive Publication Date: 2004-02-11
中国科学院发育生物学研究所
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Problems solved by technology

The former requires the pronucleus of the fertilized egg to have a certain degree of visibility (this is very difficult in quite a few animals) and the operator has a relatively high operating technique (not easy to master), and it is easy to cause nuclear damage, resulting in a very low birth rate and failure of genetic modification; The latter is unstable and poorly repeatable
However, the transfection method of in vitro cultured cells has always been troubled by the low transfection rate, and can only be used for cytological observation experiments, and cannot be used for other research.

Method used

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  • Method for transferring exogenous gene into animal cell
  • Method for transferring exogenous gene into animal cell
  • Method for transferring exogenous gene into animal cell

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Embodiment Construction

[0014] Our laboratory uses the mouse histone H2AZ (mH2AZ) as a nuclear localization signal to promote the integration of the transgenic construct bovine α-s1-casein-human G-CSF expression vector on the host chromosome, that is, the target gene expression construct and the mH2AZ gene The expression components are co-injected into the fertilized egg cytoplasm, and the target gene expression components are brought into the prokaryon by the directional migration of nuclear localization proteins. The results of this example show that the method used can bring the target gene expression components into the host cell nucleus to obtain transgene integrated mice, and this method is more convenient, practical, and easy to promote, and can bypass the bottleneck of pronuclear injection technology and avoid fertilized eggs. A large number of injuries, promote the widespread application of transgenic animal technology. Therefore, this transgenic approach can completely replace the tradition...

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Abstract

The method for transferring exogenous gene into animal cell includes the following steps: using target gene to construct target gene expression vector, using nuclear localization protein gene to construct nuclear localization protein gene expression vector and transferring the obtained target gene expression vector and nuclear localization protein gene expression vector into the cytoplasm of cultured cell or cytoplasm of animal cylula by means of cotransfection or co-injection or electroporation.

Description

technical field [0001] The invention relates to a method for transferring foreign genes into animal cells. technical background [0002] As a technical platform, transgenics can not only be used to prepare mammary gland bioreactors, but also have very important significance in disease model animals, improved animal breeds, and the study of individual development mechanisms at the molecular level. As a bioreactor, transgenic animals can produce medicinal and rare proteins that cannot be obtained by other production methods, and have the characteristics of high efficiency, low cost and more natural products. Its potential economic and social value has been valued by countries all over the world. However, the production of transgenic animals on a large scale has remained difficult for more than a decade. One of the major hurdles is the bottleneck of transgenic methods. [0003] At present, there are mainly two methods for the preparation of transgenic mammals: pronuclear mic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/64C12N15/89
Inventor 张靖溥李光三杜淼
Owner 中国科学院发育生物学研究所
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