Genetic Manipulation of Human Embryonic Stem Cells and Their Neuronal Progenies using Baculoviral Vectors

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SELECTBIO

Human embryonic stem (hES) cells have great prospect in regenerative medicine. To realize their potentials, the development of effective genetic manipulation methods in hES cells and their differentiated progenies is an obvious demand. In our lab, we demonstrated that baculoviral vectors are able to efficiently transduce hES cells and the efficiency of transient transgene expression largely depends on the transgene expression cassettes. With the incorporation of a rep gene and inverted terminal repeat sequences from adeno-associated virus, baculoviral vector can also be adapted for stable transgene expression in hES cells. These stably modified hES cells maintained the transgene expression during both self-renewal and neural differentiation process. We further studied the application of baculoviral vectors in postmitotic neurons and showed that baculoviral vectors are capable of transducing up to 80% of the hES cell-derived human neurons. With an optimized expression cassette, the transgene expression was easily observed in these baculovirus-transduced human neurons even after 3 months in vitro and at least for 4 weeks after transplantation into the brain of nude mice. Therefore, our study demonstrates that baculoviral vectors can provide an efficient tool for genetic manipulation of hES cells and their neuronal progenies.