Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 108, Issue 9, Pages 3665-3670Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1019736108
Keywords
lentivirus; vector
Categories
Funding
- Susan G. Komen for the Cure Career Catalyst Award [KG090355]
- Specialized Program of Research Excellence Developmental Grant [P50 CA058183]
- Department of Defense [BC094077]
- National Institutes of Health (NIH) [R01GM076493, R37 CA16303]
- National Institute of Environmental Health Sciences [Z01ES102745]
- US Army [W81XWH0410197]
- U.S. Department of Defense (DOD) [W81XWH0410197] Funding Source: U.S. Department of Defense (DOD)
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The discovery of RNAi has revolutionized loss-of-function genetic studies in mammalian systems. However, significant challenges still remain to fully exploit RNAi for mammalian genetics. For instance, genetic screens and in vivo studies could be broadly improved by methods that allow inducible and uniform gene expression control. To achieve this, we built the lentiviral pINDUCER series of expression vehicles for inducible RNAi in vivo. Using a multicistronic design, pINDUCER vehicles enable tracking of viral transduction and shRNA or cDNA induction in a broad spectrum of mammalian cell types in vivo. They achieve this uniform temporal, dose-dependent, and reversible control of gene expression across heterogenous cell populations via fluorescence-based quantification of reverse tet-transactivator expression. This feature allows isolation of cell populations that exhibit a potent, inducible target knockdown in vitro and in vivo that can be used in human xenotransplantation models to examine cancer drug targets.
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