Transgene pig for preparing animal model for human diseases and culturing method thereof

A technology of transgenic pigs and animal models, applied in the field of biomedicine

Inactive Publication Date: 2009-11-04
JILIN UNIV
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Problems solved by technology

However, many mouse disease models often do not fully mimic human disease

Method used

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  • Transgene pig for preparing animal model for human diseases and culturing method thereof
  • Transgene pig for preparing animal model for human diseases and culturing method thereof
  • Transgene pig for preparing animal model for human diseases and culturing method thereof

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Embodiment 1

[0040] 1. Construction of inducible expression Cre recombinase gene expression vector

[0041] 1 The important intermediate plasmids formed by plasmids are pCIneo-FRT2neoR and recombinant plasmid pMD-Cre.

[0042]2. Experimental procedure The codons of the coding sequence of the Cre recombinase gene of phage P1 were optimized according to the preference codons of the minipig gene, and the optimized Cre recombinase gene (Cre) was cloned. The porcine Mx1 promoter and BGHpolyA were respectively amplified from the pGL3Mx1 vector and the pcDNA3.1(+) vector by PCR. The FRT2NeoR cassette was obtained by digesting pGCFRT2NeoR with Xho I and SalI. The above four fragments were ligated by SOE-PCR and enzyme ligation with T4 DNA ligase, and then the Cre expression vector pET28a-Mx1-Cre-BGHpolyA-FRT2neoR was constructed using the prokaryotic expression vector PET28a (+) (see attached figure 1 ), commissioned Saibaisheng Company to synthesize SOE-PCR primers (see Table 1).

[0043] Tabl...

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Abstract

The invention provides a transgene pig for preparing an animal model for human diseases and a culturing method thereof. An inducible expression Cre recombinase expression vector pET28a-Mxl-Cre-BGHpolyA-FRT2neoR is constructed by a genetic engineering method, and can express Cre recombinase after the expression vector is induced by interferon inducer poly IC (poly I:C) so as to be used for knocking out a target gene; porcine mechanocyte capable of inducibly expressing the Cre recmbinase can be screened through technology of cell transfection and cell screening; and the pig is cloned by the technology of somatic cell nuclear transplantation. The cloned pig can be used for preparing the animal model for knocking out genes of human diseases.

Description

Technical field: [0001] The invention provides a transgenic pig for preparing an animal model of human diseases and a breeding method thereof. The transgenic pig for inducible expression of Cre recombinase used for gene knockout is prepared by somatic cell nuclear transfer technology, and belongs to the technical field of biomedicine. Background technique: [0002] Studying animal models of human diseases can help scientists understand the pathogenesis and pathological process of diseases, and even study gene therapy for diseases to cure human diseases. To date, mouse models have been widely used to study human hematopoiesis, innate and adaptive immunity, immunity, infectious diseases, cancer biology, and regenerative medicine. However, many mouse disease models often do not fully mimic human disease. Pigs are very similar to humans in terms of anatomy, physiology, organ development and disease progression. The pig animal model is the best animal model for studying human d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027
Inventor 欧阳洪生逄大欣陈立梅李莉李占军王铁东张明军任林柱闫森宋娜
Owner JILIN UNIV
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