Epididymal initial segment-specific Cre recombinase activity in Lcn8-Cre knock-in mice

Sperm acquire the ability to fertilize ova through a complex process of epididymal maturation. To identify the functions of genes expressed in the proximal epididymis, mouse models specific to this region are needed.

A Lcn8-Cre knock-in mouse line was generated using CRISPR/Cas9 technology. A 37 bp coding sequence of Lcn8 from the ATG start codon was replaced by an NLS-Cre-polyA cassette, resulting in Cre expression and the absence of Lcn8. Epididymal initial segment-specific Cre expression was identified using RT-PCR and western blotting, and the spatial-temporal Cre activity was further confirmed by using the Rosa26^tdTomato reporter mice. Immunofluorescence staining showed that active Cre recombinase was present in the principal cells. Histological analyses of sperm and epididymides, and the four-month mating tests, were used to confirm that Cre expression did not affect normal development and male fecundity.

The novel Lcn8-Cre mice can be used to establish epididymal initial segment-specific conditional knock-out mouse models.