Generation of mouse conditional knockout alleles in one step using the i-GONAD method

Successful recombination of the Fosl1 floxed allele mediated by Cre. (A) Gene structures of the Fosl1^loxP and recombined alleles following Cre-mediated DNA recombination. (B) Genotyping results of adult muscle tissues from Fosl1^loxP/loxP and littermate Fosl1^cKO mice. 5′- and 3′-loxP were detected by primers 1F + 1R and 2F + 2R, respectively. The floxed allele was detected by primers 1F + 2R. (C) Sanger sequencing result that validated DNA recombination in Fosl1^cKO muscle sample. (D) qPCR that measured Fosl1 expression in myoblasts and fibroblasts isolated from the Fosl1^cKO mice. qPCR primers are located in exon 4 which is floxed. (Ctrl) Vehicle control, (Tmx) 4-OH tamoxifen. (***) P < 0.001, (NS) not significant. Data are mean ± SEM.