动物造模,基因敲除 z-bio 2022-05-07 548

　　Objective To establish Marcksl1 knockout mice, and to preliminarily explore the effect of this gene deletion on the development of hematopoietic system.

　　Methods Marcksl1 knockout mice were constructed by CRISPR/Cas9 technology, and the mouse genotypes were identified by PCR technology and Sanger sequencing. The passage of knockout mice was analyzed by crossing knockout mice with wild-type mice. The proportion of homozygous, heterozygous and wild-type mice at different developmental stages was analyzed by crossing heterozygous mice with each other. The mouse fetal liver was isolated, and the effect of gene deletion on hematopoietic system was analyzed by blood routine and flow cytometry.

　　Results PCR combined with Sanger sequencing showed that the Marcksl1 knockout mice were successfully constructed. The genotype identification and quantitative statistics of embryonic mice at different stages showed that the embryonic death caused by the knockout of the mouse Marcksl1 gene occurred in the late embryonic development. Through blood routine and flow cytometry analysis, the results showed that Marcksl1 gene deletion did not affect the number of white blood cells, red blood cells and platelets at E15.5 d, but the proportion of hematopoietic stem cells in fetal liver increased significantly.

　　Conclusion This study successfully established Marcksl1 knockout mice, and found that the gene deletion affects the proportion of hematopoietic stem cells. This study provides an animal model for further understanding the function of this gene in embryonic development and hematopoietic system.