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Establishment of Kinase protein Akt1 conditional gene knockout mice model and application

A kinase protein, gene knockout technology, applied in the field of transgenic, can solve problems such as the inability to study the development of late embryonic organs and the regulation of glucose metabolism

Inactive Publication Date: 2012-05-30
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Since the double gene knockout mice (Akt1 / 3) died on the 12th day of embryonic development, we will not be able to study the development of vital organs and the regulation of glucose metabolism in late embryos (after 12 days)

Method used

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  • Establishment of Kinase protein Akt1 conditional gene knockout mice model and application
  • Establishment of Kinase protein Akt1 conditional gene knockout mice model and application
  • Establishment of Kinase protein Akt1 conditional gene knockout mice model and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Establishment of a conditional knockout model of the kinase Akt1 protein gene

[0055] 1. By constructing a gene targeting vector, see figure 1 ;

[0056] 2. Transfect the above vector into embryonic hepatocytes (ES), and screen the ES cell clones of homologous recombination by Southern blot hybridization, see figure 2 ;

[0057] 3. After blastocyst injection, the homologously recombined ES cells obtained in step (2) are implanted into the uterus of pseudopregnant mice, and chimeric mice are obtained after the offspring are born;

[0058] 4. Chimeric mice were further mated to obtain Akt1 knockout mice.

Embodiment 2

[0060] Establishment of a mouse model of cardiac hypertrophy in Akt1 knockout mice

[0061] The Akt1 gene-deficient mice obtained in Example 1 had no obvious abnormality at birth, and the combined knockout mice of Akt1 and Akt2 genes all died 1 month after birth; compared with wild-type control mice, the combined knockout of Akt1 and Akt2 genes In addition to the obvious thinning of the myocardial wall, loose myocardial structure, and dilated right ventricle ( Figure 4 ), suggesting that Akt1 and Akt2 gene knockout mice can be used as a model of diastolic cardiac hypertrophy.

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Abstract

The invention discloses a method for preparing a Kinase protein Akt1 conditional gene knockout non-human mammal model and application of the method. The invention further discloses a method for screening compounds for preventing or treating cardiomegaly and glucose metabolic disorders by the Kinase protein Akt1 conditional gene knockout non-human mammal model, and application of the model that Akt1 and upstream and downstream biological signal transduction pathways of the Akt1 serve as medicine intervention targets to treat cardiomegaly and glucose metabolic disorders.

Description

technical field [0001] The invention relates to the field of transgenic technology, and more specifically relates to the establishment and application of a conditional knockout animal model of the kinase protein Akt1 gene. Background technique [0002] Akt / protein kinase B (PKB) belongs to a subfamily of AGC kinase family (including PKA, PKG and PKC) - Akt / PKB family. In mammalian organisms such as mice, rats and humans, the Akt / PKB family includes three members called Akt1 / PKBα, Akt2 / PKBβ and Akt3 / PKBγ (hereinafter referred to as Akt). Akt plays an important role in the regulation of gene expression, cell survival and apoptosis, and is closely related to the occurrence and development of tumors and embryonic development. [0003] In humans and mice, these three members are encoded by three genes located on different chromosomes. The three proteins of Akt have high homology in amino acid composition, reaching more than 80%. In terms of amino acid composition, they only di...

Claims

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Application Information

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IPC IPC(8): C12N15/09A61K38/46A61P9/04A61P3/08
Inventor 杨中州常在卢双双
Owner NANJING UNIV
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