Application

• Function research of genes
• Function research of microglial cells and other Cx3cr1-expressing immune cells

Expression

Expressed gene

iCre/ERT2, improved cre recombinase and estrogen receptor 1 (human) fusion gene

Site of expression

monocytes, macrophage and brain microglia

Gene editing strategy

A targeting vector was designed to replace 390bp of exon 2 of the Cx3cr1 gene with a iCre/ERT2 (improved cre recombinase fulsed to estrogen receptor 1) coding sequence, followed by a porcine teschovirus-1 2A (P2A) and a enhanced green fluorescent protein (EGFP). The targeting vector were injected into C57BL/6N mouse zygotes. After injection, surviving 2-cell-stage zygotes were transplanted to the KM albino pseudopregnant females. The reslulting Cx3cr1-iCreERT2-EGFP founder mice were bred to C57BL/6N mice. These mice were maintained on a C57BL/6N background.

Phenotype Analysis

The expression of EGFP and tdTomato in mouse spleen, bone marrow, and brain mononuclear macrophages was detected by flow cytometry, and Cx3cr1 (Mut /+) was found after induction of Tamoxifen.The majority of macrophages in the brain of tdTomato (Mut/+) mice were double positive cells of EGFP and tdTomato, while the mononuclear macrophages in spleen and bone marrow were single positive cells of EGFP. The results were consistent with the results in the study, indicating that B-Cx3cr1-iCreERT2-EGFP mice were designed and expressed successfully.

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2. YonaSimon,Kim Ki-Wook,Wolf Yochai et al. Fate mapping reveals origins and dynamics of monocytes and tissue macrophages under homeostasis.[J] .Immunity, 2013, 38: 79-91. [PMID: 23273845]

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4. Ushijima T , Fujimoto N , Matsuyama S , et al. The actin-organizing formin protein Fhod3 is required for postnatal development and functional maintenance of the adult heart in mice[J]. Journal of Biological Chemistry, 2017:jbc.M117.813931. [PMID: 29158260]

5. Madisen L ,Zwingman T A , Sunkin S M , et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain[J]. NATURE NEUROSCIENCE, 2009, 13(1):133-140. [PMID: 20023653]

Microglial FoxO3a deficiency ameliorates ferroptosis-induced brain injury of intracerebral haemorrhage via regulating autophagy and heme oxygenase-1.pdf

When using the B-Cx3cr1-iCreERT2-EGFP mouse strain in a publication, please indicate the originate from Biocytogen and stock #110779 in your Materials and Methods section.