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The common carp ( Cyprinus carpio ) as one of the most important aquaculture fishes produces over 3 million metric tones annually, approximately 10% the annual production of the all farmed freshwater fish worldwide. However, the tetraploidy genome and long generation-time of the common carp have made its breeding and genetic studies extremely difficult. Here, TALEN and CRISPR-Cas9, two versatile genome-editing tools, are employed to target common carp bone-related genes  sp7, runx2, bmp2a, spp1, opg , and muscle suppressor gene  mstn . TALEN were shown to induce mutations in the target coding sites of  sp7, runx2, spp1  and  mstn . With CRISPR-Cas9, the two common carp  sp7  genes,  sp7a  and  sp7b , were mutated individually, all resulting in severe bone defects; while  mstnba  mutated fish have grown significantly more muscle cells. We also employed CRISPR-Cas9 to generate double mutant fish of  sp7a;mstnba  with high efficiencies in a single step. These results demonstrate that both TALEN and CRISPR-Cas9 are highly efficient tools for modifying the common carp genome, and open avenues for facilitating common carp genetic studies and breeding.

Targeted disruption of sp7 and myostatin with CRISPR-Cas9 results in severe bone defects and more muscular cells in common carp