Lent-On-Plus lentiviral vectors for conditional expression in human stem cells
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URI: https://hdl.handle.net/10481/99312Metadatos
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2016-11Resumen
Conditional transgene expression in human stem cells has been difficult to achieve due to the low
efficiency of existing delivery methods, the strong silencing of the transgenes and the toxicity of the
regulators. Most of the existing technologies are based on stem cells clones expressing appropriate
levels of tTA or rtTA transactivators (based on the TetR-VP16 chimeras). In the present study, we aim
the generation of Tet-On all-in-one lentiviral vectors (LVs) that tightly regulate transgene expression
in human stem cells using the original TetR repressor. By using appropriate promoter combinations
and shielding the LVs with the Is2 insulator, we have constructed the Lent-On-Plus Tet-On system
that achieved efficient transgene regulation in human multipotent and pluripotent stem cells. The
generation of inducible stem cell lines with the Lent-ON-Plus LVs did not require selection or cloning,
and transgene regulation was maintained after long-term cultured and upon differentiation toward
different lineages. To our knowledge, Lent-On-Plus is the first all-in-one vector system that tightly
regulates transgene expression in bulk populations of human pluripotent stem cells and its progeny.