title: The pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo.
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.
authors.affiliation: Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: Kristen L
authors.initials: KL
authors.lastname: Meerbrey
authors.forename: Guang
authors.initials: G
authors.lastname: Hu
authors.forename: Jessica D
authors.initials: JD
authors.lastname: Kessler
authors.forename: Kevin
authors.initials: K
authors.lastname: Roarty
authors.forename: Mamie Z
authors.initials: MZ
authors.lastname: Li
authors.forename: Justin E
authors.initials: JE
authors.lastname: Fang
authors.forename: Jason I
authors.initials: JI
authors.lastname: Herschkowitz
authors.forename: Anna E
authors.initials: AE
authors.lastname: Burrows
authors.forename: Alberto
authors.initials: A
authors.lastname: Ciccia
authors.forename: Tingting
authors.initials: T
authors.lastname: Sun
authors.forename: Earlene M
authors.initials: EM
authors.lastname: Schmitt
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Bernardi
authors.forename: Xiaoyong
authors.initials: X
authors.lastname: Fu
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Bland
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Cooper
authors.forename: Rachel
authors.initials: R
authors.lastname: Schiff
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Rosen
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Westbrook
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Elledge
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000473:pathology; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D003952:Diagnostic Imaging; D005260:Female; D015870:Gene Expression; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D049449:Luminescence; D008321:Mammary Glands, Animal / Q000378:metabolism / Q000473:pathology; D051379:Mice; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D015203:Reproducibility of Results; D023041:Xenograft Model Antitumor Assays
chemical_list: D018076:DNA, Complementary; D034741:RNA, Small Interfering