Interference of transgene mouse model by slow viral vectors mediation IKK alpha gene RNA and use thereof

A technology of transgenic mice and α gene, which is applied in the field of medicine and medical treatment, and can solve problems such as irreplaceable

Inactive Publication Date: 2008-05-28
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY +1
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  • Application Information

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Problems solved by technology

However, in vitro research work cannot replace the actual situation in vivo.

Method used

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  • Interference of transgene mouse model by slow viral vectors mediation IKK alpha gene RNA and use thereof
  • Interference of transgene mouse model by slow viral vectors mediation IKK alpha gene RNA and use thereof
  • Interference of transgene mouse model by slow viral vectors mediation IKK alpha gene RNA and use thereof

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Experimental program
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Effect test

Embodiment

[0092] Example: Preparation method of lentiviral vector-mediated IKKα gene RNA interference transgenic mice

[0093] 1. Synthesis of two oligonucleotide chains.

[0094] According to the online RNAi sequence design software of Invitrogen Company of the United States, search for the sequence of the knock-down IKKα gene, and synthesize two oligonucleotide chains, the sequences of which are as follows:

[0095] 5′-CACCGCAGGCTCTTTCAGGGACACGAATGTCCCTGAAAGAGCCTGC-3′

[0096] 5′-AAAAGCAGGCTCTTTCAGGGACATTCGTGTCCCTGAAAGAGCCTGC-3′

[0097] An annealing reaction was then carried out, and the annealed product was named IKKαds Oligo, and the structure after annealing was as follows:

[0098]

[0099] 2. Preparation of expression vector (pU6-IKKα-knockdown expression vector).

[0100]The above IKKαds Oligo was ligated with 2 μl pENT / U6vector (0.5ng / μl) to transform TOP 10 competent bacteria, ligated with 2 μl pENT / U6vector (0.5ng / μl) to transform TOP10 competent bacteria, and then the...

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Abstract

The invention discloses an IKK alpha gene RNA Interfere mouse model mediated by lentiviral vector and usage. The experiment constructs the lentiviral vector aiming at IKK alpha gene RNAi (knockdown), the vector is inducted into perivitelline space of fertilized eggs of a mouse to obtain transgenic mouse model which carries the vector, and the average positive rate of founder mice is 15% via the verification of a PCR. Then detection of a reverse transcription PCR and a method of real-time quantitative PCR shows that the expression level of peripheral blood cells IKK alpha mRNA of the transgenic mice is obviously decreased. The biological function of IKK alpha gene and the relation between the abnormal expression and tumor can be researched in vivo by employing the model, thereby finding anti-tumor drugs.

Description

technical field [0001] The present invention relates to the field of medicine and medical technology, in particular to the use of transgenic mouse models for tumor research, and more specifically to a biological method for the establishment of IKKα gene RNAi transgenic mouse models and the study of IKKα genes in vivo. The biological function and its application in relation to the occurrence and development of tumors. Therefore, the above model provides a good foundation for a more comprehensive and in-depth understanding of the regulatory pathway of IKKα-NF-κB in the future and for its use in disease research. Background technique [0002] (A) NF-κB protein family. [0003] The NF-κB protein family plays a very important role in normal metabolism and immune response. Usually, the NF-κB protein is combined with the inhibitory protein Iκ-Bs in the cytoplasm under physiological conditions. External stimuli cause phosphorylation of Iκ-B protein, which is then degraded by prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/86A01K67/027
Inventor 杨志峰王铸钢毛积芳
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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