RT Journal Article
T1 Lent-On-Plus Lentiviral vectors for conditional expression in human stem cells.
A1 Benabdellah, Karim
A1 Muñoz, Pilar
A1 Cobo, Marién
A1 Gutierrez-Guerrero, Alejandra
A1 Sánchez-Hernández, Sabina
A1 Garcia-Perez, Angélica
A1 Anderson, Per
A1 Carrillo-Gálvez, Ana Belén
A1 Toscano, Miguel G
A1 Martin, Francisco
AB 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.
YR 2016
FD 2016-11-17
LK http://hdl.handle.net/10668/10609
UL http://hdl.handle.net/10668/10609
LA en
DS RISalud
RD Apr 7, 2025