The PINDUCER Lentiviral Toolkit for Inducible RNA Interference in Vitro and in Vivo

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2011 Feb 11

PMID 21307310

Citations 435

Authors

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

Ronald J Bernardi

Xiaoyong Fu

Christopher S Bland

Thomas A Cooper

Rachel Schiff

Jeffrey M Rosen

Thomas F Westbrook

Stephen J Elledge

Affiliations

Soon will be listed here.

Abstract

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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