Breeding and genotype identification of CCR2 knockout mice
10.19405/j.cnki.issn1000-1492.2025.07.001
- Author:
Huiru Zhang
1
;
Anqi Wang
1
;
Chong Liu
1
;
Yuanyuan Zhou
1
;
Hui Xue
1
;
Jiajie Tu
1
Author Information
1. Institute of Clinical Pharmacology，School of Pharmaceutical Sciences，Anhui Medical University，Hefei 230032
- Publication Type:Journal Article
- Keywords:
C-C motif chemokine receptor type 2;
knockout;
polymerase chain reaction;
genotyping;
protein expression;
rheumatoid arthritis
- From:
Acta Universitatis Medicinalis Anhui
2025;60(7):1167-1172
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To explore the breeding and genotyping of CCR2 knockout mice, and to verify the applicability of the polymerase chain reaction(PCR) method for genotype detection of CCR2 knockout mice.
Methods: The introduced CCR2 pure male mice and wild-type female mice were mated and bred to produce the offspring generation, the obtained F1 generation heterozygous mice were continued to be mated. DNA was extracted by clipping the tail tissues of the mice at the age of 2 weeks, the target gene fragment was amplified by PCR, and the genotypic results were determined by agarose gel electrophoresis. The proportion of purebred progeny carrying the CCR2 knockout gene was increased by genetic crosses, the effect of CCR2 knockout in the progeny mice was verified by using Western blot against major immune cells and key organs, and flow cytometry was used to detect whether the knockout of the CCR2 gene had any effect on the function of the immune system by targeting the major immune cells.
Results:CCR2 knockout mice were successfully bred and characterized, and three genotypes of F2 generation mice were obtained: CCR2+/+, CCR2+/-, and CCR2-/-. The offspring genotypes were identified by PCR, and Western blot showed extremely low CCR2 protein expression in CCR2 knockout mice. Flow analysis showed that CCR2 knockdown reduced the expression of CD4+T and Th1 cells in mouse spleen-derived T cells, but did not affect macrophage function.
Conclusion :Correct breeding and identification are important ways to get the pure CCR2 knockout mice, and PCR method for identifying mouse genotypes is simple, fast and reliable.
- Full text:2026030417082692488CCR2基因敲除小鼠的构建及基因型鉴定_张慧茹.pdf
