Generation of Fbxl13 Large Deletion Mice

Fbxl13 large deletion (LD) KO mice were generated as previously described [33 (link)]. We designed two gRNAs that recognize either exon 1 or exon 7 to avoid the cleavage of nearby genes (S2A Fig) and inserted the sequences into the pX459 V2.0 plasmid (#62988, Addgene, Cambridge, MA, USA). EGR-G01 embryonic stem (ES) cells [34 (link)] were co-transfected with 1.0 μg of two gRNAs inserted vectors. Nine ES clones out of 48 had a large deletion of Fbxl13. ES cell clones that possessed the mutation were injected into ICR embryos and chimeric blastocysts were transferred into the uteri of pseudopregnant females. Generated chimeric male mice were mated with B6D2F1 female mice to obtain heterozygous KO mice. Fbxl13 KO mice were maintained by sibling matings.