Morula complementation restores male germline in NANOS2 null sheep
The spermatogonial-specific gene NANOS2 was disrupted in male (NANOS2+/-, NANOS2-/-) and female (NANOS2-/-) ovine fetal fibroblasts via gRNA-Cas9-mediated homology-directed repair. Targeted cell strains and wild-type controls were used to produce cloned offspring for breeding and phenotyping. Male monoallelic and female biallelic targeting of NANOS2 did not affect germline development, resulting in fertile animals capable of producing fertile offspring with normal reproductive performance.