Infectious and parasitic diseases have major negative economic and animal welfare impacts on aquaculture of salmonid species. Improved knowledge of the functional basis of host response and genetic resistance to these diseases is key to developing preventative and treatment options. Cell lines provide valuable models to study infectious diseases in salmonids, and genome editing using CRISPR/Cas systems provides an exciting avenue to evaluate the function of specific genes in those systems. While CRISPR/Cas editing has been successfully performed in a Chinook salmon cell line (CHSE-214), there are no reports to date of editing of cell lines derived from the most commercially relevant salmonid species Atlantic salmon and rainbow trout, which are difficult to transduce and therefore edit using lentivirus-mediated methods. In the current study, a method of genome editing of salmonid cell lines using ribonucleoprotein (RNP) complexes was optimised and tested in the most commonly used salmonid fish cell lines: Atlantic salmon (SHK-1 and ASK cell lines), rainbow trout (RTG-2) and Chinook salmon (CHSE-214). Electroporation of RNP based on either Cas9 or Cas12a was efficient at targeted editing of all the tested lines (typically > 90% cells edited), and the choice of enzyme expands the number of potential target sites for editing within the genomes of these species. These optimised protocols will facilitate functional genetic studies in salmonid cell lines, which are widely used as model systems for infectious diseases in aquaculture.

传染病和寄生虫病对鲑鱼物种的水产养殖产生重大的负面经济和动物福利影响。更好地了解宿主对这些疾病的反应和遗传抗性的功能基础是开发预防和治疗选择的关键。细胞系为研究鲑鱼中的传染病提供了有价值的模型，而使用CRISPR / Cas系统进行基因组编辑则为评估这些系统中特定基因的功能提供了令人兴奋的途径。尽管已经在Chinook鲑鱼细胞系（CHSE-214）中成功进行了CRISPR / Cas编辑，但迄今为止尚无关于编辑最商业相关鲑鱼物种大西洋鲑和虹鳟鱼的细胞系的报道，这些很难转导并因此使用慢病毒介导的方法进行编辑。在目前的研究中 优化了一种使用核糖核蛋白（RNP）复合物对鲑鱼细胞系进行基因组编辑的方法，并在最常用的鲑鱼鱼细胞系中进行了测试，该方法包括：大西洋鲑鱼（SHK-1和ASK细胞系），虹鳟鱼（RTG-2）和奇努克族三文鱼（CHSE-214）。基于Cas9或Cas12a的RNP的电穿孔可有效靶向编辑所有受试品系（通常> 90％的细胞已被编辑），酶的选择扩大了这些物种基因组中潜在靶标位点的数量。这些优化的方案将促进鲑鱼细胞系的功能遗传研究，鲑鱼细胞系被广泛用作水产养殖传染病的模型系统。