CRISPR/Cas9-mediated knock-in of the murine Y chromosomal genes Zfy1 and Zfy2.

ABSTRACT:

Background

The Y-linked mouse zinc finger genes Zfy1 and Zfy2 are critical fertility factors in mice but the mechanisms by which they regulate spermatogenesis remain unclear. We recently produced Zfy1/2 double knock-out mice and observed a complete loss in fertility. However, the biochemical mechanism by which Zfy regulates spermatogenesis is unknown, and ZFY expression has not yet been confirmed at the protein level. As both Zfy homologues share ~ 95% sequence similarity, it is difficult to produce an anti-ZFY antibody specific to either homologue.

Results

To overcome this technical challenge, we used CRISPR/Cas9 genome editing to develop tagged Zfy1 knock-in (XY^Zfy1-HA), Zfy2 knock-in (XY^Zfy2-FLAG, XY^Zfy2-3xFLAG, and XY^Zfy2-HA),

SUBMITTER: Holmlund H 

PROVIDER: S-EPMC12831244 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature