Establishment of cell-based transposon-mediated transgenesis in cattle.

ALESSIO A; FILI A; GARRELS W; FORCATO D; OLMOS NICOTRA MF; LIAUDAT AC; BEVACQUA R; SAVY V; HIRIART MI; TALLURI TR; OWENS JB; IVICS Z; SALAMONE D; MOISYADI S; KUES W; BOSCH P

THERIOGENOLOGY

ELSEVIER SCIENCE INC

Lugar: Amsterdam; Año: 2015

0093-691X

Transposon-mediated transgenesis is a well-established tool for genome modification insmall animal models. However, translation of this active transgenic method to large animalswarrants further investigations. Here, the piggyBac (PB) and sleeping beauty (SB)transposon systems were assessed for stable gene transfer into the cattle genome. Bovinefibroblasts were transfected either with a helper-independent PB system or a binary SBsystem. Both transposons were highly active in bovine cells increasing the efficiency ofDNA integration up to 88 times over basal nonfacilitated integrations in a colony formationassay. SB transposase catalyzed multiplex transgene integrations in fibroblast cells transfectedwith the helper vector and two donor vectors carrying different transgenes (fluorophoreand neomycin resistance). Stably transfected fibroblasts were used for SCNT andon in vitro embryo culture, morphologically normal blastocysts that expressed the fluorophorewere obtained with both transposon systems. The data indicate that transpositionis a feasible approach for genetic engineering in the cattle genome.