Fig. 2
Cx3cr1-CreERT2 rat genetic characterization. (A) The Cx3cr1-CreERT2 rat was created from a bacterial artificial chromosome (BAC) containing the rat Cx3cr1 gene (CHORI) and recombineered to with the coding region of CreERT2 and the poly-adenylation signal of bovine growth hormone (bGHpA) immediately after the start codon of the rat Cx3cr1 gene. The primer positions (black arrows) for PCR genotyping of the 5’ and 3’ junctions of the CreERT2 DNA cassette. The positions of the primers (blue arrows) and probe (green arrow) for a droplet digital PCR (ddPCR)- and digital PCR (dPCR)- based genotyping assay/gene expression assays are shown along the bottom. (B) Digital gel image showing PCR results from the 5’ and 3’ junction assays in non-carrier animals (WT), hemizygous (Hem), and homozygous (Hom) carriers of the Cx3cr1-CreERT2 transgene. The 5’ and 3’ junction amplicons are 550 bp and 1553 bp, respectively. (C) ddPCR assay results from genomic DNA isolated from hemizygous and homozygous animals. Relative RNA expression of Cx3cr1 locus (D) and Cx3cr1-CreERT2 locus in WT and transgenic animals using dPCR. As expected, only transgenic animals express the CreERT2 transgene (t-test p < 0.001). Partially created in BioRender. Harvey, B. (2025) https://BioRender.com/95l7337.
