动物造模,基因敲除小鼠模型 z-bio 2021-02-22 316

　　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 for preliminary phenotyping analysis. The sgRNA expression plasmid was constructed by designing a single guide RNA sgRNA recognition sequence according to the base sequence of the first exon of activin α subunit. 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 mutation of the inhibin α subunit gene of newborn mice was detected by PCR and gene sequencing. Select homozygous male and female mice for F2 gene knockout, collect male testes and female ovaries for observation, and perform paraffin sections and HE staining.

　　Result: A total of 12 FO generation activin 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 proportion of each genotype in the F2 generation of 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 activin gene knockout mouse model. The cancerous phenotype of F2 homozygous mice indicates that activin plays an important role in the mouse reproductive system.