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Construction method of zebra fish animal model for cardiovascular disease drug screening

An animal model and construction method technology, applied in the fields of botanical equipment and methods, compound screening/testing, biochemical equipment and methods, etc., can solve the problem of difficulty in obtaining zebrafish mutants with cardiovascular malformations, etc. Easy to obtain, small size effect

Active Publication Date: 2020-07-31
华芯微鱼(苏州)生物科技有限公司
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Problems solved by technology

[0004] However, in actual use, such as in the process of operation, it is difficult to obtain genetically stable cardiovascular malformation zebrafish mutants

Method used

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  • Construction method of zebra fish animal model for cardiovascular disease drug screening
  • Construction method of zebra fish animal model for cardiovascular disease drug screening
  • Construction method of zebra fish animal model for cardiovascular disease drug screening

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Embodiment Construction

[0038] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.

[0039] The present invention provides a method for constructing a zebrafish animal model for cardiovascular disease drug screening. The specific steps are as follows:

[0040] (1) Selection of gRNA target site of zebrafish heg1 gene and in vitro synthesis of gRNA: To obtain the target target, first download the genome data of the target gene heg1 from the zebrafish genome database, and select the 529bp long target near the first exon of heg1 gene Enter the sequence in...

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Abstract

The invention discloses a construction method of a zebra fish animal model for cardiovascular disease drug screening, and specifically relates to the field of animal model construction methods. The method comprises the following specific steps: selecting a gRNA target site of a zebra fish heg1 gene and synthesizing gRNA in vitro; in order to obtain the target site, firstly, downloading genome dataof the target gene heg1 from a zebra fish genome database; and selecting a 529bp long target sequence near a first exon of the heg1 gene and inputting the sequence into a target point website to obtain a target point primer. According to the invention, a cardiovascular malformation zebra fish mutant with genetic stability can be obtained; by using the zebra fish embryo transparency and the autonomous oxygen dissolving property, the mutant can be used for screening and verifying the influence of different cardiovascular drugs on cardiovascular development; and the zebra fish animal model construction method provides important physiological significance and specificity for screening of cardiovascular disease treatment drugs.

Description

Technical field [0001] The invention relates to the field of methods for constructing animal models, in particular to a method for constructing a zebrafish animal model for cardiovascular disease drug screening. Background technique [0002] Heg1 (heart of glass) gene is a transmembrane receptor. It is an embryonic recessive lethal mutation identified by John D.Mably in the zebrafish ENU screening in 2003. It has specific atrial and ventricular enlargement and blood flow retardation. Cardiovascular disease phenotype, Heg1 mainly regulates the adhesion between the heart and blood vessel cells. Studies in mice have shown that heg1 gene mutations can weaken the endothelial barrier, leading to sepsis, atherosclerosis and multiple sclerosis. In vascular diseases, the homologous gene of heg1 can be detected in humans, mice and rats, which makes it possible to apply the experimental data obtained from the heg1 mutant to human clinical practice. [0003] CRISPR (Clustered regularly inters...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/90C12N15/89C12N15/12A01K67/027A61K49/00
CPCC12N15/902C12N15/89C07K14/705A01K67/0276A61K49/0008A01K2217/075A01K2227/40A01K2267/0375Y02A40/81
Inventor 田静卢淑娴张东伟
Owner 华芯微鱼(苏州)生物科技有限公司
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