Fig. 1.
Strategy for creating tendon targeted Col11a1 conditional knockout mice. (A) Schematic diagram showing the wild-type allele, targeting vector, targeted allele, and floxed allele as well as the excised allele after Cre recombination of Col11a1 gene. The Col11a1 targeting vector was obtained from the KOMP Repository (project ID: CSD80258). Exon 3 of Col11a1 is flanked with LoxP sites and the Neo cassette is flanked with FRT sites. Location of primers for LacZ, as well as 3’ and 5’ primers used to determine insert orientation are shown (red arrows). The large blue and green arrows indicate the location of the primers used to identify specific alleles. (B) Strategy for gene targeting of embryonic stem cells and generation of tendon-targeted Col11a1 conditional-knockout mice. Linearized Col11a1 targeting vector was electroporeated into ES cells. Blastocysts were injected with targeted ES cells yielding chimeric mice. The chimeric mice were crossed with wild type mice yielding F1 progeny with the targeted allele (Col11a1+/ta). Breeding of Col11a1+/ta with FLPe mice resulted in excision of FRT flanked sequences, removal of the Neo cassette and creation of conditional knockout mice (Col11a1flox/flox). These mice were bred with mice expressing ScxCre to target the knockout to tendons (Col11a1Δten/Δten).