Construction and gene identification of CSF1R +/-mice
10.19405/j.cnki.issn1000-1492.2025.05.015
- Author:
Yuanyuan Zhou
1
;
Chong Liu
1
;
Anqi Wang
1
;
Huiru Zhang
1
;
Jiaqi Qiu
1
;
Mengjuan Zhu
1
;
Jiajie Tu
1
Author Information
1. School of Pharmaceutical Sciences , Anhui Medical University , Hefei 230032)
- Publication Type:Journal Article
- From:
Acta Universitatis Medicinalis Anhui
2025;60(5):884-889
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To constructCSF1R+/-mice and to analyze their genotypes, so as to provide animal model basis for disease pathological mechanism and drug target.
Methods : A linearized targeting vector was designed according to Cre/Loxp system. A Loxp site was inserted upstream of the 5th exon of theCSF1Rgene, and a neomycin resistance box with Loxp sites on both sides was inserted downstream of the 5th exon. The linearized targeting vector was electroporated into embryonic stem cells. The correctly targeted embryonic stem cells were injected into the blastocysts of C57BL/6J mice to obtain chimeric mice, which were bred with Zp3-Cre mice. The newborn mice were numbered 9-14 days after birth and their tails were cut. The DNA of the mice was extracted, and the genotype of the mice was identified by polymerase chain reaction and agarose gel electrophoresis. The expression of CSF1R in mouse macrophages was detected by flow cytometry. The expression of CSF1R in mouse tissues was detected by Western blot.
Results: The results of agarose gel electrophoresis showed that 453 bp bands were amplified in wild type mice, and 453 bp and 650 bp bands were amplified in heterozygous mice. The results of flow cytometry showed that the expression of CSF1R in peritoneal macrophages and bone marrow-derived macrophages of CSF1R heterozygous mice was lower than that of WT group(P<0.05). The results of Western blot showed that the expression of CSF1R in spleen, kidney and brain tissue of CSF1R heterozygous group was lower than that of WT group(P<0.05).
Conclusion:CSF1R+/-mice are successfully constructed, reproduced and identified, which provides an animal model basis for further revealing the potential mechanism of CSF1R in immune regulation.
- Full text:2026020515233970848CSF1R~(+_-)小鼠的构建和基因鉴定_周园园.pdf
