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Transgenic animal over-expressing cardiac ankyrin repeat protein as well as production method and application thereof

A technique for transgenic animals and repeat sequences, applied to transgenic animals overexpressing myocardial anchored repeat sequence proteins, its production and application fields

Inactive Publication Date: 2011-01-26
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Zou et al found in cardiomyocytes that CARP can inhibit the expression of the TK promoter fused with the HF-1 sequence, but has no effect on the expression of the basic TK promoter

Method used

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  • Transgenic animal over-expressing cardiac ankyrin repeat protein as well as production method and application thereof
  • Transgenic animal over-expressing cardiac ankyrin repeat protein as well as production method and application thereof
  • Transgenic animal over-expressing cardiac ankyrin repeat protein as well as production method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Production and Identification of CARP Transgenic Mice

[0034] 1.1 Generation of CARP transgenic mice

[0035] The 960bp mouse CARP cDNA was connected to the promoter of the heart-specific myosin heavy chain (α-MHC), the myc tag indicated the expression of exogenous CARP protein, and the hGH polyA site was used to guide the correct transcription termination, so Constructed expression vectors such as figure 1 As shown, after the correctness of the clone was confirmed by DNA sequencing, the clone was named α-MHC-CARP. The α-MHC promoter can guide the specific expression of downstream genes in cardiomyocytes. Except for the transient expression at embryonic day 9.5-10.5, it is mainly expressed in the ventricular myocardium after birth and continues until adulthood.

[0036] The purified α-MHC-CARP linear DNA was microinjected into the male pronucleus of mouse fertilized eggs, transferred to pseudopregnant mice, and the offspring of the mice were identified. O...

Embodiment 2

[0055] Example 2. CARP transgenic mice partially inhibit ISO-induced cardiac hypertrophy

[0056] Male CARP-Tg and WT mice aged 9-11 weeks were selected and randomly divided into four groups: WT-sham, WT-ISO, Tg-sham, Tg-ISO. Under anesthesia, a mini-pump containing ISO solution was transplanted into the back of the mouse, at a dose of 30 mg / kg / d, to induce cardiac hypertrophy in the mouse heart; mice in the control group were added with ascorbic acid solution. Fourteen days after Pump transplantation, the changes of myocardial hypertrophy in the mice were observed by echocardiography, and the mice were sacrificed under anesthesia, and the degree of myocardial hypertrophy was judged by HW / BW and HW / TL indicators.

[0057] 2.1 CARP transgenic mice can partially inhibit the increase in heart volume induced by isoproterenol

[0058] After 14 days of ISO induction, the heart of WT mice showed myocardial hypertrophy, the ratio of HW / BW and HW / TL increased, the volume of the heart ...

Embodiment 3

[0078] Example 3. CARP transgenic mice partially inhibit TAC-induced cardiac hypertrophy

[0079] 3.1 TAC-induced increase in mouse heart volume was partially alleviated in CARP transgenic mice

[0080] After 4 weeks of TAC induction, the heart of wild-type mice showed significant cardiac hypertrophy. From the appearance of the mouse heart, it can be found that the mouse heart volume increased (such as Figure 8 B); Longitudinal section of mouse heart tissue (such as Figure 8 F) or cross-cut slices (such as Figure 8 J) Analysis also showed that after TAC induction, the heart volume of the mice increased, the ventricular wall became thicker, etc., showing obvious concentric myocardial hypertrophy.

[0081] However, in CARP-Tg mice, after 4 weeks of TAC induction, there was no significant increase in heart volume ( Figure 8 D, H), the trend of ventricular wall hypertrophy was also partially suppressed ( Figure 8 L), showing that CARP transgenic mice can partially inhibit...

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Abstract

The invention relates to a transgenic animal over-expressing a cardiac ankyrin repeat protein as well as a production method and application thereof. The transgenic animal specifically over-expresses the cardiac ankyrin repeat protein in a heart tissue can partially inhibit the cardiac hypertrophy induced by isoprenaline and / or the cardiac hypertrophy induced by heat aortic coarctation.

Description

technical field [0001] The present invention relates to a non-human transgenic animal overexpressing Cardiac Ankyrin Repeat Protein (CARP) and its production method and application. The transgenic animal specifically overexpresses the CARP gene in heart tissue and can partially inhibit isoproterenol Cardiac hypertrophy induced by sodium glucocorticoid (ISO) and cardiac aortic constriction (transverse aortic constriction, TAC) induced cardiac hypertrophy. Background technique [0002] Cardiac hypertrophy is a complication of common clinical diseases such as hypertension, valvular disease, acute myocardial infarction and congenital heart disease. Myocardial hypertrophy or cardiac hypertrophy refers to the increase in the volume and weight of the ventricle, which is an adaptive response of cardiomyocytes to external mechanical stimulation or nerve-humoral stimulation, which can increase cardiac output and pump blood function in a short period of time. The body adapts to change...

Claims

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Application Information

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IPC IPC(8): A01K67/027C12N15/85A61K38/17A61P9/00
Inventor 许家林李艳凤朱大海
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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