id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
work_h2mcv33dbng4dgnjwyh4eblewi	Wiebke Garrels	One-step Multiplex Transgenesis via Sleeping Beauty Transposition in Cattle	2016	6	.pdf	application/pdf	4161	399	54	Pablo Bosch4, Kerstin Pötzsch5, Esther Grueso5, Zoltán Ivics5 & Wilfried A. genome editing and genetic engineering based on somatic cell nuclear transfer (SCNT) suffer from a low overall efficiency1–4, due to extensive micromanipulation and nuclear reprogramming failures. Here, we established a highly efficient one-step gene transfer system into the bovine genome of in vitro fertilized (IVF) zygotes, allowing the simultaneous integration of several independent transgenes delivered by the two (pT2CAGGS-Venus and pT2Cryaa-tdTomato) or three transposons (pT2CAGGS-Venus, pT2CryaatdTomato and pT2Casein-E2) were co-injected into the opaque cytoplasm of a bovine zygote. absence of helper plasmid and transposon backbone sequences in the genomes of the transgenic calves was confirmed by PCR (Fig. 1h). transfer after one week of in vitro culture likely contributed to the 100% rate of multi-transgenic calves per born fertilized zygotes, and embryo transfer of IVF-derived blastocysts typically result in pregnancy rates of no 	./cache/work_h2mcv33dbng4dgnjwyh4eblewi.pdf	./txt/work_h2mcv33dbng4dgnjwyh4eblewi.txt