<> "The repository administrator has not yet configured an RDF license."^^ . <> . . "The development of a novel S/MAR DNA vector platform for the stable, persistent and safe genetic engineering of dividing cells"^^ . "The development of episomally maintained DNA vectors that are capable of providing safe,\r\npersistent and stable modification of cells whilst avoiding the risk of integration-mediated\r\ngenotoxicity would provide a valuable tool for genetic research.\r\nDNA vectors harboring a Scaffold/Matrix Attachment Region (S/MAR) can provide persistent and\r\nrobust transgene expression in human cancer cell lines which can be used in in vitro, in vivo and ex\r\nvivo studies. A prototype S/MAR DNA vector with which we initiated this study replicates\r\nepisomally, remains unsilenced and unmethylated following the genetic modification of cells.\r\nAlthough, it showed great promise it does have significant limitations which restricts its application.\r\nThe establishment rate of the original DNA vector is an inefficient passive process and the selection\r\nprocedure is lengthy and often produces drug-resistant but non-expressing colonies generating a\r\nmosaic of cells with differential transgene expression. Thus, these vectors represent a reasonable\r\ntool for simple studies such as cell labelling with reporter genes but are not suitable for more\r\nsophisticated work such as gene-rescue experiments or for the genetic engineering of primary\r\nhuman cells. In this project, we have refined and enhanced the S/MAR DNA vector system. The\r\nrange of next-generation DNA vectors that we have produced provide several advances over the\r\noriginal vectors. We have demonstrated that this new S/MAR DNA vector platform is more\r\nefficient and stable with improved efficiency in establishing stable cell-lines. We have also\r\ndemonstrated that the persistence of transgene expression and the molecular integrity of the\r\nvector has been improved in a range of cancer cell lines as well as in primary human cells.\r\nWe have used this next-generation of DNA vector to generate labelled cells suitable for in vitro and\r\nin vivo drug screening. We have also generated isogenic tumor models which provide insights into\r\nthe mechanism of pancreatic cancer development by restoring crucial tumor suppressor genes to\r\nthe cells without altering the molecular or biochemical integrity of the cells. Additionally, we have\r\nutilized the vector system to persistently modify a range of dividing cell types including primary\r\nmouse embryonic stem cells and embryonic fibroblasts and primary human fibroblasts.\r\nAs an ultimate demonstration of the efficacy of this DNA vector we have used it to genetically\r\nmodify human T-cells for immunotherapy and have demonstrated it to be capable of expressing\r\ntransgenes in these cells for over 1 month with minimal toxicity.\r\nWe have demonstrated that this novel class of DNA vector can be used to persistently modify\r\nevery cell-type tested providing sustained and high levels of transgene expression whilst avoiding\r\nthe risk of insertional mutagenesis induced by the random integration of genetic material with\r\nminimal impact to the cell."^^ . "2018" . . . . . . . "Matthias"^^ . "Bozza"^^ . "Matthias Bozza"^^ . . . . . . "The development of a novel S/MAR DNA vector platform for the stable, persistent and safe genetic engineering of dividing cells (PDF)"^^ . . . "Matthias Bozza Thesis.pdf"^^ . . . "The development of a novel S/MAR DNA vector platform for the stable, persistent and safe genetic engineering of dividing cells (Other)"^^ . . . . . . "lightbox.jpg"^^ . . . "The development of a novel S/MAR DNA vector platform for the stable, persistent and safe genetic engineering of dividing cells (Other)"^^ . . . . . . "preview.jpg"^^ . . . "The development of a novel S/MAR DNA vector platform for the stable, persistent and safe genetic engineering of dividing cells (Other)"^^ . . . . . . "medium.jpg"^^ . . . "The development of a novel S/MAR DNA vector platform for the stable, persistent and safe genetic engineering of dividing cells (Other)"^^ . . . . . . "small.jpg"^^ . . . "The development of a novel S/MAR DNA vector platform for the stable, persistent and safe genetic engineering of dividing cells (Other)"^^ . . . . . . "indexcodes.txt"^^ . . "HTML Summary of #23821 \n\nThe development of a novel S/MAR DNA vector platform for the stable, persistent and safe genetic engineering of dividing cells\n\n" . "text/html" . . . "420 Englisch"@de . "420 English"@en . . . "600 Technik, Medizin, angewandte Wissenschaften"@de . "600 Technology (Applied sciences)"@en . . . "610 Medizin"@de . "610 Medical sciences Medicine"@en . .