title: Transcriptionally regulated and nontoxic delivery of the hyperactive Sleeping Beauty Transposase.
abstract: The Sleeping Beauty (SB) transposase and, in particular, its hyperactive variant SB100X raises increasing interest for gene therapy application, including genome modification and, more recently, induced pluripotent stem cells (iPS) reprogramming. The documented cytotoxicity of the transposase, when constitutively expressed by an integrating retroviral vector (iRV), has been circumvented by the transient delivery of SB100X using retroviral mRNA transfer. In this study, we developed an alternative, safe, and efficient transposase delivery system based on a tetracycline-ON regulated expression cassette and the rtTA2(S)-M2 transactivator gene transiently delivered by integration-defective lentiviral vectors (IDLVs). Compared with iRV-mediated delivery, expression of tetracycline-induced SB100X delivered by an IDLV results in more efficient integration of a GFP transposon and reduced toxicity. Tightly regulated expression and reactivation of the transposase was achieved in HeLa cells as wells as in human primary keratinocytes. Based on these properties, the regulated transposase-IDLV vectors may represent a valuable tool for genetic engineering and therapeutic gene transfer.
authors: Department of Life Sciences, Centre for Regenerative Medicine, University of Modena and Reggio Emilia , Modena, Italy.
authors: Fabienne
authors: F
authors: Cocchiarella
authors: Department of Life Sciences, Centre for Regenerative Medicine, University of Modena and Reggio Emilia , Modena, Italy.
authors: Maria Carmela
authors: MC
authors: Latella
authors: Department of Life Sciences, University of Modena and Reggio Emilia , Modena, Italy.
authors: Valentina
authors: V
authors: Basile
authors: Department of Life Sciences, Centre for Regenerative Medicine, University of Modena and Reggio Emilia , Modena, Italy.
authors: Francesca
authors: F
authors: Miselli
authors: Institute of Experimental Hematology, Hannover Medical School , Hannover, Germany.
authors: Melanie
authors: M
authors: Galla
authors: Department of Life Sciences, University of Modena and Reggio Emilia , Modena, Italy.
authors: Carol
authors: C
authors: Imbriano
authors: Department of Life Sciences, Centre for Regenerative Medicine, University of Modena and Reggio Emilia , Modena, Italy.
authors: Alessandra
authors: A
authors: Recchia