Novel Hyperactive Transposons for Genetic Modification of Induced Pluripotent and Adult Stem Cells: A Nonviral Paradigm for Coaxed Differentiation

作者：Belay Eyayu; Matrai Janka; Acosta Sanchez Abel; Ma Ling; Quattrocelli Mattia; Mates Lajos; Sancho Bru Pau; Geraerts Martine; Yan Bing; Vermeesch Joris; Rincon Melvin Yesid; Samara Kuko Ermira; Ivics Zoltan; Verfaillie Catherine; Sampaolesi Maurilio; Izsvak Zsuzsanna; VandenDriessche Thierry; Chuah Marinee K L^*

来源：Stem Cells, 2010, 28(10): 1760-1771.

DOI：10.1002/stem.501

• Transposon
• Sleeping Beauty
• iPS
• Myoblast
• Stem cell
• Mesenchymal stem cell
• Muscle

Adult stem cells and induced pluripotent stem cells (iPS) hold great promise for regenerative medicine. The development of robust nonviral approaches for stem cell gene transfer would facilitate functional studies and potential clinical applications. We have previously generated hyperactive transposases derived from Sleeping Beauty, using an in vitro molecular evolution and selection paradigm. We now demonstrate that these hyperactive transposases resulted in superior gene transfer efficiencies and expression in mesenchymal and muscle stem/progenitor cells, consistent with higher expression levels of therapeutically relevant proteins including coagulation factor IX. Their differentiation potential and karyotype was not affected. Moreover, stable transposition could also be achieved in iPS, which retained their ability to differentiate along neuronal, cardiac, and hepatic lineages without causing cytogenetic abnormalities. Most importantly, transposon-mediated delivery of the myogenic PAX3 transcription factor into iPS coaxed their differentiation into MYOD+ myogenic progenitors and multinucleated myofibers, suggesting that PAX3 may serve as a myogenic "molecular switch" in iPS. Hence, this hyperactive transposon system represents an attractive nonviral gene transfer platform with broad implications for regenerative medicine, cell and gene therapy. STEM CELLS 2010; 28: 1760-1771

• 出版日期2010-10