Journal
GENE THERAPY
Volume 17, Issue 1, Pages 14-25Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/gt.2009.109
Keywords
humanized mouse model; inducible gene expression; lentiviral vector; hematopoietic stem cell; Tet-On system
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Funding
- Dutch AIDS Foundation [2005022]
- International AIDS Vaccine Initiative (IAVI)
- Technology Foundation STW
- NIH/NIAD [R21-AI073136]
- Marie Curie Intra-European fellowship [MEIF-CT-2007-039689]
- Bill and Melinda Gates Foundation
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [R21AI073136] Funding Source: NIH RePORTER
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The efficient control of gene expression in vivo from lentiviral vectors remains technically challenging. To analyze inducible gene expression in a human setting, we generated 'human immune system' (HIS) mice by transplanting newborn BALB/c Rag2(-/-) IL-2R gamma(-/-)(c) immunodeficient mice with human hematopoietic stem cells transduced with a doxycycline-inducible lentiviral vector. We compared several methods of doxycycline delivery to mice, and could accurately measure doxycycline in vivo using a new sensitive detection assay. Two different lentiviral vector designs with constitutive (TRECMV-V14) or autoregulatory (TREAuto-V14) expression of an optimized reverse tetracycline transactivator were used to transduce human hematopoietic stem cells. After transplantation into immunodeficient mice, we analyzed the expression of the green fluorescent protein (GFP) reporter gene in the human hematopoiesis-derived cells that develop and accumulate in the generated HIS mice. We show efficient inducible GFP expression in adult HIS mice containing TREAuto-V14-transduced human cells, whereas GFP expression is poor with the TRECMV-V14 vector. Multiple cycles of doxycycline exposure in the TREAuto-V14 group result in repeated cycles of GFP expression with no loss of intensity. These findings are of major interest for gene therapy and basic research settings that require inducible gene expression. Gene Therapy (2010) 17, 14-25; doi: 10.1038/gt.2009.109; published online 3 September 2009
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