抑制素基因敲除小鼠模型,动物造模 z-bio 2021-10-29 555

　　Purpose: Use clustered, regularly spaced short-interval palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)] gene editing technology to construct an inhibin knockout mouse model, and perform preliminary phenotypic analysis to create one Types of.

　　``By designing a single guide RNA sgRNA recognition sequence based on the base sequence of the first exon of the inhibin α subunit according to the method, an sgRNA expression plasmid was constructed. After transcribing sgRNA and Cas9 mRNA in vitro with T7NA polymerase, the sgRNA/Cas9 mRNA was microinjected into the fertilized eggs of C57BL/6J mice, and the gene bases of inhibin α subunits of newborn mice were detected by PCR and gene sequencing. mutation. Male and female mice with homozygous F2 gene knockout were selected, male testes and female mice ovaries were observed, and paraffin sections and HE staining analysis were performed.

　　Result: A total of 12 FO generation inhibin knockout mice were obtained. Eight male mice and C57BL/6J female mice were backcrossed to obtain F1 generation mice, and then mated to obtain F2 generation mice. The genotype ratio of F2 generation mice conforms to Mendel's law, and knockout mice are not embryonic lethal. F2 homozygous mice developed cancerous testes and ovaries and were unable to give birth.

　　Conclusion: We have successfully constructed an inhibin gene knockout mouse model. The cancerous phenotype of F2 homozygous mice indicates that inhibin plays an important role in the mouse reproductive system.