Enzymatic engineering of the porcine genome with transposons and recombinases

摘要: Swine is an important agricultural commodity and biomedical model. Manipulation of the pig genome provides opportunity to improve production efficiency, enhance disease resistance, add value swine products. Genetic engineering can also expand utility pigs for modeling human disease, developing clinical treatment methodologies, or donating tissues xenotransplantation. Realizing full potential genetic requires translation complete repertoire tools currently employed in smaller model organisms practical use pigs. Application transposon recombinase technologies manipulation characterization their activity cells. We tested four systems- Sleeping Beauty, Tol2, piggyBac, Passport cultured porcine Transposons increased efficiency DNA integration up 28-fold above background provided precise delivery 1 15 transgenes per cell. Both Cre Flp were functional cells as measured by ability remove a positive-negative selection cassette from 16 independent clones over 20 genomic locations. demonstrated Cre-dependent switch capable eliminating intervening activating GFP expression episomal genome-resident transposons. have first time that transposons recombinases are mobilizing into out efficient manner. This study basis based genome.