Introduction : Organoids are in vitro models derived from stem cells that recapitulate many aspects of the complex structure and function of the corresponding in vivo tissue. We previously characterized organoid cultures from different segments of the porcine gut vs. the original tissue by RNA-seq (data not shown). A vast majority of expressed genes was shared between organoids and their tissue counterparts. Innate immune and epithelial gene lists maintained a higher proportion of shared genes vs. the full gene catalogue, confirming that organoids are a good in vitro “3Rs” proxy to evaluate genotype-to-phenotype relationships involving innate immunity responses of the gut epithelia. These systems could be more appropriate than regular cell lines for host-virus interaction studies. Aim: We aim to optimize cutting-edge genome-editing tools and protocols on pig intestinal organoids. As a proof of concept, we generate knock-out organoids for the aminopeptidase N (ANPEP) gene, a well-known receptor of the transmissible gastroenteritis coronavirus (TGEV). We report here preliminary results on the transfection efficiency of porcine gut organoids by electroporation, adapting the method described for human organoids (Fujii et al, 2015) and by a GFP-lentiviral vector transduction.