Salt Sensitive Tet-Off-Like Systems to Knockdown Primordial Germ Cell Genes for Repressible Transgenic Sterilization in Channel Catfish, Ictalurus Punctatus

Repressible knockdown approaches were investigated for transgenic sterilization in channel catfish, . Two primordial germ cell (PGC) marker genes, and , were targeted for knockdown, and an off-target gene, , was monitored. Two potentially salt sensitive repressible promoters, zebrafish (ADSS) and zebrafish (Rm), were each coupled with four knockdown strategies: ds-sh RNA targeting the 5' end (N1) or 3' end (N2) of channel catfish , full-length cDNA sequence of channel catfish for overexpression (cDNA) and ds-sh RNA targeting channel catfish (DND). Each construct had an untreated group and treated group with sodium chloride as the repressor compound. Spawning rates of full-sibling P₁ fish exposed or not exposed to the constructs as treated and untreated embryos were 93% and 59%, respectively, indicating potential sterilization of fish and repression of the constructs. Although the mRNA expression data of PGC marker genes were inconsistent in P₁ fish, most F₁ individuals were able to downregulate the target genes in untreated groups and repress the knockdown process in treated groups. The results indicate that repressible transgenic sterilization is feasible for reproductive control of fish, but more data from F₂ or F₃ are needed for evaluation.