Genetically modified pigs produced with a nonviral episomal vector

Abstract

Genetic modification of cells and animals is an invaluable tool

Persistent episomal transgene expression in liver following delivery of a scaffold/matrix attachment region containing non-viral vector

“…Impressively, an S/MAR-containing nonviral vector has been recently used to develop genetically modified pigs. 27 An S/MAR element has also been incorporated, among other elements, into lentiviral vectors 28 and into a plasmid vector already optimized for long-term expression. 13 The mechanism by which S/ MAR elements provide the unique ability to sustain long-term transgene expression in vivo has, however, so far not been specifically investigated.…”

Evaluation of swine fertilisation medium (SFM) efficiency in preserving spermatozoa quality during long-term storage in comparison to four commercial swine extenders

“…For this reason, there has been increased interest in developing new extenders and in establishing the optimal storage conditions for diluted spermatozoa. In the last few decades, we utilised a homemade diluent (swine fertilisation medium (SFM)) for swine spermatozoa manipulation and biotechnological applications as the production of transgenic pigs utilising the sperm-mediated gene transfer technique (Lavitrano et al, 2002;Lavitrano et al, 2003), with good fertility results as well as in AI or surgical or laparoscopic inseminations Manzini et al, 2006). Nevertheless, this SFM has never been evaluated methodically in comparison to other well-defined commercial diluents.…”

Insulin expressed from endogenously active glucose-responsive EGR1 promoter in bone marrow mesenchymal stromal cells as diabetes therapy

“…Recent advances in vector engineering such as the construction of human artificial chromosomes 45 and inclusion of a scaffold matrix attachment region in plasmid DNA for stable episomal retention reportedly achieve durable transgene expression even in the face of active cell proliferation and could be vectors of choice for BMMSC. 22,47 Notwithstanding the fact that only about half of normal insulin secretion was reconstituted in our study, this level of insulin restoration is clinically meaningful as life-threatening diabetic ketoacidosis is prevented in type I diabetics with merely 10% of physiological circulating insulin. 48 However, it is evident that improvements in reconstituting insulin secretion beyond what we have achieved in this study are needed for a more robust therapeutic effect.…”