动物造模,基因敲除 z-bio 2022-05-16 294

　　OBJECTIVE: To breed and identify FOXO3A knockout mice. The selected FOXO3A knockout heterozygous mice were used for breeding, and the homozygous mice were used for experiments. Preliminary analysis of the effect of FOXO3A knockout on hematopoietic cells The effect of FOXO3A gene on hematopoietic cell injury will lay a foundation for the subsequent research on the regulation of hematopoietic cell injury.

　　Methods: Heterozygous female mice were mated with wild-type male mice, and heterozygous female mice and heterozygous male mice were selected and bred to obtain homozygous mice by one male and one female or one male and two females in the same cage. ;The bred young mice were marked by clipping their toes, and their tails were clipped to extract genomic DNA. PCR was used to identify the genotype. Two bands of 100bp/186bp were identified as heterozygous mice (FOXO3A+/-), and a band of 186bp was identified. The homozygous mice (FOXO3A-/-) were identified, and the wild-type mice (FOXO3A+/+, WT) were identified to obtain a 100bp band; flow cytometry was used to analyze the identified wild-type and homozygous mice Bone marrow cell typing was performed.

　　RESULTS: FOXO3A knockout mice have been successfully bred and identified. The offspring of homozygous knockout mice and heterozygous mice grow normally, and there is no significant difference in appearance, growth and development compared with wild-type FVB/N mice. ; A certain number of homozygous mice have been obtained for subsequent experiments; the results of bone marrow cell count showed that the bone marrow nucleated cell count of FOXO3A knockout homozygous mice was compared with that of wild-type mice [(6.167±1.424)vs .(10±1.732)], no significant difference (P=0.1625); flow analysis results showed that the proportion of hematopoietic progenitor cells (lin-scal-1-ckit+, HPC) in the bone marrow of FOXO3A knockout mice was significantly increased (P<0.05), there="" was="" no="" significant="" difference="" in="" the="" proportion="" of="" hematopoietic="" stem="" cells="" p="">0.05).

　　Conclusion: FOXO3A knockout mice have been successfully obtained by genotype identification. Preliminary investigation of FOXO3A knockout has no significant effect on bone marrow cell count and typing in mice, and the effect of FOXO3A knockout on mouse hematopoietic cells needs further research Experiments prove it.