Nonviral Gene Delivery with the Sleeping Beauty Transposon System

Abstract

Effective gene therapy requires robust delivery of therapeutic genes into relevant target cells, long-term gene expression, and minimal risks of secondary effects. Nonviral gene transfer approaches typically result in only short-lived transgene expression in primary cells, because of the lack of nuclear maintenance of the vector over several rounds of cell division. The development of efficient and safe nonviral vectors armed with an integrating feature would thus greatly facilitate clinical gene therapy studies. The latest generation transposon technology based on the Sleeping Beauty (SB) transposon may potentially overcome some of these limitations. SB was shown to provide efficient stable gene transfer and sustained transgene expression in primary cell types, including human hematopoietic progenitors, mesenchymal stem cells, muscle stem/progenitor cells (myoblasts), induced pluripotent stem cells, and T cells. These cells are relevant targets for stem cell biology, regenerative medicine, and gene-and cell-based therapies of complex genetic diseases. Moreover, the first-in-human clinical trial has been launched to use redirected T cells engineered with SB for gene therapy of B cell lymphoma. We discuss aspects of cellular delivery of the SB transposon system, transgene expression provided by integrated transposon vectors, target site selection of the transposon vectors, and potential risks associated with random genomic insertion.

Sleeping Beauty Transposition

“…The first clinical application of the SB system will help to validate both the safety and efficacy of this approach. Applications of the SB system fall outside the scope of this chapter, and readers are referred to recent review articles (160,161,162,163,164,165,166,167,168,169,170,171,172).…”

Periostin Secreted by Mesenchymal Stem Cells Supports Tendon Formation in an Ectopic Mouse Model

“…Up to 900 genes may be modified by lentiviral infection of CD34 + cells [32]. These problems ask for special safety regards before lentiviral vectors can proceed to the clinic [33] or, for example, the use of nonviral gene delivery systems, such as the ''sleeping beauty,'' may be advisable [34]. In spite of all these caveats, it has been documented that in vitro and even in vivo lentiviral infection of MSCs did not affect their multilineage differentiation potential [35] and in our case, tenogenic tissue formation of control implantations with GFP-lentiviruses or uninfected MSCs was not observed (data not shown).…”

Sleeping Beautytransposition: from biology to applications

“…By exploiting the local hopping feature of the SB system, it was possible to decipher the genomic neighborhood of a particular integration site by remobilizing the transposons from individual chromosomes (Keng et al, 2005;Kokubu et al, 2009). The greatest success of SB has been in its use for phenotype-driven genetics (Carlson et al, 2003;Elso et al, 2015;Horie et al, 2003;Ivics & Izsvak, 2011;Izsvak et al, 2010;Moriarity & Largaespada, 2011). Unlike a candidate-based strategy, SB transposon-based gene trapping, enhancer trapping and unbiased mutagenesis approach is suitable to identify genes with unexpected phenotypes (Balciunas et al, 2004;Lu et al, 2007;Song & Cui, 2013).…”