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Introgression of farmed salmon escapees into wild stocks is a major threat to the genetic integrity of wild populations. Using germ cell-free fish in aquaculture may mitigate this problem. Our study investigated whether it is possible to produce germ cell-free salmon in F0 by using CRISPR-Cas9 to knock out  dnd , a factor required for germ cell survival in vertebrates. To avoid studying mosaic animals, sgRNA targeting  alb  was simultaneously used as a visual tracer since the phenotype of  alb  KO is complete loss of pigmentation. Induced mutations for the tracer ( alb ) and the target ( dnd ) genes were highly correlated and produced germ cell-less fish lacking pigmentation, underlining the suitability of  alb  KO to serve as tracer for targeted double allelic mutations in F0 animals in species with prohibitively long generation times. This is also the first report describing  dnd  knockout in any fish species. Analyzing gene expression and histology of  dnd  KO fish revealed that sex differentiation of the somatic compartment does not depend on the presence of germ cells. However, the organization of the ovarian somatic compartment seems compromised in mutant fish.

Dnd knockout ablates germ cells and demonstrates germ cell independent sex differentiation in Atlantic salmon