Open AccessThe pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo
Author(s) -
Kristen L. Meerbrey,
Guang Hu,
Jessica D. Kessler,
Kevin Roarty,
Mamie Z. Li,
Justin E. Fang,
Jason I. Herschkowitz,
Anna E. Burrows,
Alberto Ciccia,
Tingting Sun,
Earlene M. Schmitt,
R Bernardi,
Xiaoyong Fu,
Christopher S. Bland,
Thomas A. Cooper,
Rachel Schiff,
Jeffrey M. Rosen,
Thomas F. Westbrook,
Stephen J. Elledge
Publication year - 2011
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.1019736108
Subject(s) - rna interference , gene knockdown , biology , computational biology , small hairpin rna , transduction (biophysics) , transactivation , gene expression , in vivo , gene silencing , microbiology and biotechnology , gene , genetics , rna , biochemistry
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.