title: Lent-On-Plus Lentiviral vectors for conditional expression in human stem cells.
abstract: 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.
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.|LentiStem Biotech. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Karim
authors.initials: K
authors.lastname: Benabdellah
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Muñoz
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Marién
authors.initials: M
authors.lastname: Cobo
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Gutierrez-Guerrero
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Sabina
authors.initials: S
authors.lastname: Sánchez-Hernández
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Angélica
authors.initials: A
authors.lastname: Garcia-Perez
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Per
authors.initials: P
authors.lastname: Anderson
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Ana Belén
authors.initials: AB
authors.lastname: Carrillo-Gálvez
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Miguel G
authors.initials: MG
authors.lastname: Toscano
authors.affiliation: Genomic Medicine Department. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.|LentiStem Biotech. GENYO, Centre for Genomics and Oncological Research, Pfizer-University of Granada-Andalusian Regional Goverment, Parque Tecnológico Ciencias de la Salud, Av. de la Ilustración 114, 18016 Granada, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Martin
mesh_terms: D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D014161:Transduction, Genetic