Method, primer and plasmid for constructing EPO gene knockout zebrafish animal model and preparation method of primer and plasmid

Abstract

The invention provides a method, primer and plasmid for constructing an EPO gene knockout zebrafish animal model and a preparation method of the primer and the plasmid, and belongs to the field of biomedicine. The method includes the steps of 1) establishment of an EPO gene knockout zebrafish CRISPR oligomer sequence plasmid; and 2) establishment and culture of the EPO gene knockout zebrafish animal model on the basis of CRISPR gene knockout technology; wherein the step 1) comprises 1-1) design and synthesis of the primer aiming at an EPO exon 2 region target sequence and 1-2) synthesis of the Escherichia coli plasmid containing the EPO target sequence. The method for constructing the EPO gene knockout zebrafish animal model can be used to construct the EPO gene knockout zebrafish animal model, and facilitates further understanding and research of EPO.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for constructing an EPO gene knockout zebrafish animal model based on CRISPR gene knockout technology, primers, plasmids and a plasmid preparation method. Background technique [0002] Erythropoietin (EPO) is a hormone-like substance secreted by the kidney and liver, which plays an important role in inhibiting the apoptosis of primitive erythrocytes and promoting erythropoiesis. Recent studies have shown that EPO plays an important regulatory role in the occurrence and development of heart, kidney, liver diseases, diabetes, hyperlipidemia and other metabolic syndromes, and plays a significant role in oxidative stress and anti-apoptosis. [0003] Due to the low success rate of gene knockout and complicated steps in the preparation of EPO gene knockout based on zinc lipoprotein, Talen and other gene editing technologies, and because the role of EPO is impor...

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