Targeted Gene Insertion and the Effect of Random Insertion of the Masou Salmon Delta5-Desaturase Transgene on n-3 Fatty Acid Production and Culture Traits of Channel Catfish (Ictalurus punctatus)
Type of DegreeMaster's Thesis
Fisheries and Allied Aquacultures
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Channel catfish (Ictalurus punctatus) is one of the most abundant catfish species in North America. The objectives of this study were to determine the effectiveness of the Masou salmon delta5-desaturase transgene driven by a beta-actin promoter for improving n-3 fatty acid production in F1 transgenic channel catfish, Ictalurus punctatus together with proximate analysis, as well as to determine pleiotropic effects of disease resistance. Additionally, a novel approach of using multiple transgenes to alter multiple steps in a pathway, which has not been previously reported in aquaculture species was utilized. As a result, Apo14-Masousalmon_Elovl2 gene was successfully inserted on chromosome 1 in one channel catfish utilizing microinjection and CRISPR/Cas9 mediated knockin. Transgenic fish showed a 33% increase in n-3 fatty acid level, and a 15% decrease in n-6 fatty acids level. Masou salmon delta5-desaturase transgene driven by a beta-actin promoter affected n-3 fatty acid production in a positive direction. Insertion of the masou salmon delta5-desaturase transgene into channel catfish had major effects on metabolism, physiology, body composition, and growth of channel catfish. A major goal of increasing the percentage of n-3 fatty acids was achieved, but the objective of increasing total n-3 fatty acids was not. Body composition was altered, pleiotropic effects on performance traits were observed and the transgene appeared to increase uniformity of these traits.