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Method for searching ALK3 downstream gene related to congenital heart disease-ventricular septal defect

A technology for congenital heart disease and ventricular septal defect, which is applied in the field of searching for genes related to disease-causing diseases, and can solve problems such as difficult screening of downstream genes and unclear organ specificity

Inactive Publication Date: 2010-05-12
杨德业
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Problems solved by technology

Therefore, it can be determined that ALK3 has a great relationship with cardiac ventricular septal defect, and ALK3 is a relatively upstream signaling molecule related to ventricular septal defect, but unfortunately it is still unclear about organ specificity, especially heart specificity. The downstream genes of ALK3 are more resistant, and the main reason is that the downstream genes are difficult to screen, which is also a major problem in international research at present.

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  • Method for searching ALK3 downstream gene related to congenital heart disease-ventricular septal defect
  • Method for searching ALK3 downstream gene related to congenital heart disease-ventricular septal defect
  • Method for searching ALK3 downstream gene related to congenital heart disease-ventricular septal defect

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Embodiment Construction

[0026] The present invention is specifically described below by the embodiment, only for further illustrating the present invention, can not be interpreted as the limitation of protection scope of the present invention, the technical engineer of this field can make some non-essential improvements and improvements to the present invention according to the content of the above-mentioned invention Adjustment.

[0027] (1) Obtaining of heart-specific ALK3 gene knockout mice: mating of female α-MHC Cre+ / -, ALK3+ / - and male ALK3F / F (the fragment of Lox P inserted on both sides of the second exon of ALK3 gene) α-MHC Cre+ / -, ALK3F / - and α-MHC Cre+ / -, ALK3F / + mouse embryos can be obtained. Such as figure 1 As shown, wherein, the black rectangle represents the ALK3 gene, the white rectangle represents the knockout of the ALK3 gene, and the black arrow represents the loxP sequence.

[0028] (2), extraction of total RNA and mRNA: adopt total RNA and mRNA kit to extract and purify total ...

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Abstract

The invention discloses a method for searching an ALK3 downstream gene related to congenital heart disease-ventricular spetal defect. The method comprises the following steps: adopting rat embryo (11.5 days) of Alpha-MHC-Cre+ / -, ALK3F / -(testing group) and Alpha-MHC-Cre+ / -, ALK3F / +(contrasting group), and using a PCR selective cDNA difference display method and a gene chip method to carry out initial screening, thus obtaining possible ALK3 downstream candidate genes; subsequently adopting an RT-PCR method with lower cost to eliminate false positive ALK3 downstream candidate genes; subsequently using an in-situ hybrid method to carry out expression part validation on the ALK3 downstream candidate genes remained after the elimination by the RT-PCR method; and approving the ALK3 downstream gene related to be the congenital heart disease-ventricular septal defect. The method provides a theoretical base for developing the curative gene medicine of ventricular septal defect and finds new curative targets for exploring new medicine.

Description

technical field [0001] The invention relates to a method for searching for genes related to disease pathogenesis, in particular to a method for searching for ALK3 downstream genes related to the pathogenesis of congenital heart disease-ventricular septal defect. Background technique [0002] Congenital heart disease is mainly caused by obstacles in fetal heart development. In recent years, it has become one of the hotspots to reveal the pathogenesis of congenital heart disease through the study of heart development. Precisely positioning abnormal genes as gene therapy has become a congenital heart disease. Treatment has laid the groundwork, but the underlying causes of cardiac developmental disorders, especially the pathogenesis of ventricular septal defects, are still not fully understood. According to foreign research in recent years, transgenic rat models knocking out one or several genes are constantly being established, among which Chisaka and Capecchi established a rat...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 杨德业
Owner 杨德业
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