Transgenic Tumor Animal Models

Summary

Animal models of transgene tumor are constructed by taking the genes that produce certain tumors as exogenous genes, which can help people to study the activity of oncogenes, the mechanism of tumor development and the process of tumor development, and provide experimental basis for the treatment of tumors. At present, the construction of transgenic tumor animal models mainly adopts the following routes: directly transfer oncogenes and genes promoting tumorigenesis and development into the animal body, so that such genes can be expressed in specific tissues or non-specific expression; transfer of tumor angiogenesis regulation genes, from the gene level to regulate tumor angiogenesis, the use of tumor vascular growth factors to promote or inhibit tumor formation and growth; transfer of tumor metastasis promotion genes, mainly metallo-matrix proteases; transfected tumor cell drug resistance related genes, etc.

Operation method

Preparation of transgenic tumor-bearing animals

Materials and Instruments

Equipment:
Micromanipulator

Move

Preparation of transgenic tumor animals is done in the following steps depending on the type of receptor:

(i) Microinjection: A

. The target gene constructed in vitro is injected directly into the prokaryotic nucleus of a fertilized egg in the prokaryotic period using a micromanipulator with a very fine micropipette, and then it is implanted into a pseudopregnant female mouse.

B. Integration of an exogenous gene into the genome when the recipient embryo is undergoing DNA synthesis or repair.

(B) Retroviral infection: A

. Highly concentrated viral particles are made by inserting exogenous DNA into the genome of a retrovirus, and exogenous genes are integrated into the genome of the recipient by viral infection of early embryonic cells;

B, transgenic animals can be obtained by injecting vector viruses into the blastocyst cavity;

C, It is also possible to directly co-culture embryos with monolayer cultured cells capable of releasing retroviruses.

(C) Embryonic stem cell-mediated: A

, Exogenous genes are usually introduced into stem cells using liposome-mediated, electroshock, or retroviral infection

;

B, The host embryo at the blastocyst stage is then transferred, and finally the host embryo is transplanted into the uterus of a pseudopregnant female rat;

C. Transgenic animals mediated by embryonic stem cells can then be obtained.

(D) Sperm vector: A

. Mature spermatozoa are pre-cultured with exogenous DNA;

B. By methods such as electroporation or liposome mediation, the spermatozoa are given the ability to carry exogenous DNA and enter the egg to fertilize it;

C. Integration of exogenous DNA into chromosomes.

(E) Somatic cell nuclear transplantation: A

. Transfect animal somatic cells with exogenous target genes by liposome-mediated methods, and select somatic cells with exogenous target genes as nuclear donors.

B. Transplantation of donor nuclei into denucleated imprinted cells.

(F) Gene knockout:A. According to the principle of homologous recombination, the exogenous DNA is transferred into the cell to recombine with a specific homologous sequence on the endogenous chromosome, so as to achieve the purpose of genome modification.

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