Editing the Melanocortin-4 Receptor Gene in Channel Catfish Using the CRISPR-Cas9 System

The melanocortin-4 receptor (MC4R) plays a critical role in homeostasis and the regulation of body weight. Polymorphisms in the mc4r gene have been discovered and linked to growth, carcass composition, and meat quality traits. Therefore, we used the CRISPR-Cas9 system to target the mc4r gene in the most important freshwater aquaculture species in the USA, channel catfish, Ictalurus punctatus. Guide RNAs were designed to direct the Cas9 to the coding sequence of the channel catfish mc4r gene. gRNA(s)-Cas9 mixtures were delivered into one-cell embryos using electroporation and microinjection. For each treatment, the nature and rate of mutations were analyzed. Hatching and survival rates were calculated. The overall mutation rates were 30.6% and 66.7-90.6% for electroporation and microinjection, respectively. Mutated fish generated via electroporation or microinjection exhibited 38% and 20% improvement in body weight, respectively, when compared with the full-sib control. The mean feed conversion ratio of the mutants was 1.18 compared with 1.57 in the control fish. The improved growth and feed conversion indicate that the generation of mc4r-edited fish could economically benefit aquaculture production.