Novel hyperactive transposons for genetic modification of induced pluripotent and adult stem cells: a nonviral paradigm for coaxed differentiation.
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.
Belay
Eyayu
E
Flanders Institute for Biotechnology (VIB), Vesalius Research Center, University of Leuven, Leuven, Belgium.
Mátrai
Janka
J
Acosta-Sanchez
Abel
A
Ma
Ling
L
Quattrocelli
Mattia
M
Mátés
Lajos
L
Sancho-Bru
Pau
P
Geraerts
Martine
M
Yan
Bing
B
Vermeesch
Joris
J
Rincón
Melvin Yesid
MY
Samara-Kuko
Ermira
E
Ivics
Zoltán
Z
Verfaillie
Catherine
C
Sampaolesi
Maurilio
M
Izsvák
Zsuzsanna
Z
Vandendriessche
Thierry
T
Chuah
Marinee K L
MK
D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism*; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D032342:Muscle Cells / Q000166:cytology / Q000378:metabolism; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D018626:Retroelements / Q000235:genetics / Q000502:physiology*; D019895:Transposases / Q000378:metabolism
D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D018626:Retroelements; C068686:Pax3 protein, mouse; D019895:Transposases