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A rapid and scalable system for studying gene function in mice using conditional RNA interference

ABSTRACT: Four libraries were analyzed to assess the level and effects of inducing the ectopic expression of a luciferase targeting miRNA in mouse embryonic fibroblasts (MEFs). Here, we find low level, leaky expression in the absence of ROSA-rtTA and doxycyclin. When expression is effectively induced, the Luciferase miRNA is expressed as the 8th most abundant species in the cells. Keyword(s): Epigenetics

ORGANISM(S): Mus musculus

PROVIDER: GSE19476 | GEO | 2010/12/01

SECONDARY ACCESSION(S): PRJNA122263

REPOSITORIES: GEO

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A rapid and scalable system for studying gene function in mice using conditional RNA interference

Project description:Four libraries were analyzed to assess the level and effects of inducing the ectopic expression of a luciferase targeting miRNA in mouse embryonic fibroblasts (MEFs). Here, we find low level, leaky expression in the absence of ROSA-rtTA and doxycyclin. When expression is effectively induced, the Luciferase miRNA is expressed as the 8th most abundant species in the cells. Keyword(s): Epigenetics 4 libraries were analyzed

2010-12-01 | E-GEOD-19476 | biostudies-arrayexpress

Single cell RNA-seq of Gif lineage cells in homeostasis and upon injury

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Effect of Hyp, low phosphate diet and sex on mRNA gene expression in the mouse kidney

Project description:The renal adaptation to changing dietary phosphate levels is not well understood. The dominant Hyp mutation of the Phex gene in mice causes X-linked hypophosphatemia with low renal retention of phosphate and blocks physiologic adaptation to low phosphate diets. At P < 0.01, there were 1,711 transcripts significantly affected by genotype, 1,428 by diet and 5,601 by sex. Many renal transporters other than phosphate, as well as many novel transcripts of unknown function, were affected by the Hyp mutation. Some genes for fat metabolism and inflammation were up-regulated in Hyp kidneys. Of the genes affected by genotype and diet, only 378 were affected by both. In summary, the Hyp mutation induced changes in mRNA levels for numerous transcripts exceeding that required to alter phosphate retention. The data suggest broader physiological roles for the Phex gene unrelated to phosphate conservation. Keyword = Phex Keyword = Hyp Keyword = mouse Keyword = kidney Keyword = sex Keyword = mRNA Keyword = microarray Keyword = low phosphorus diet Keywords: disease state analysis

2007-01-01 | GSE5243 | GEO

SAGE Profiles of Rat Lung Tissues

Project description:Thirty-two SD rats (male, 150 grams, 50days) were randomly divided into four groups: normal control group, asthma group, acupuncture-treated asthma group and acupuncture-treated normal group. The rat lungs were removed immediately after the rats were died. The rat lungs were conserved in liquid nitrogen. SAGE analyses were dependent on the classic protocol (Velculescu VE, et al, Serial Analysis of Gene Expression Detailed Protocol, version 1.0e, 2000) available at the SAGENET website. Keyword = Rat Keyword = Lung Keyword = Asthma Keyword = SAGE Keyword = Acupuncture Keywords: disease state analysis

2005-04-09 | GSE2475 | GEO

Expression data from primary mammary epithelial cells expressing ectopic p190B RhoGAP

Project description:P190B RhoGAP is required for mammary gland development, and its overexpression disrupts mammary gland branching morphogenesis. To better understand the mechanisms by which p190B regulates mammary gland development we performed gene expression microarray analysis on mammary epithelial cells isolated from p190B overexpressing transgenic mice compared to control mice. The mice used in this study were previously developed to inducibly overexpress p190B selectively in the mammary gland (Vargo-Gogola 2006). FVB/N mice carrying the MMTV-rtTA and TetO-p190B-IRES-luciferase transgenes or the MMTV-rtTA transgene only (Gunther EJ, FASEB) were fed doxycycline (Dox) chow (2 g/kg) for 7 d prior to removal of mammary glands and isolation of primary mammary epithelial cells at 6 weeks of age.

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Specific MicroRNAs Are Preferentially Expressed by Skin Stem Cells To Balance Self-Renewal and Early Lineage Commitment

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2011-01-04 | E-GEOD-26396 | biostudies-arrayexpress

SAGE Profiles of Rat Lung Tissues

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In-vivo inhibition of Fgfr2b signalling in embryonic whole mouse lungs at E14.5

Project description:In this experiment we inhibited FGF signalling in E14.5 mouse lungs by using a dominant negative mouse model. We are interested in assessing the genetic regulation by FGF signalling at this stage of lung development. Specifically, we used a tetracycline controlled transcriptional activation model (Tet(o)) to manipulate FGF signalling. Genetically modified male mice that expressed a reverse tetracycline-controlled trans activator (rtTA) on the Rosa26 locus and a soluble FGFR2b on the Tet(o) promoter (Tet(o)sFgfr2b) (B6.Cg-Gt(ROSA)26Sortm1.1(rtTA,EGFP)NagyTg(tetO-Fgfr2b/Igh)1.3Jaw/sbel), were crossed with females lacking the Tet(o)sFgfr2b allele. Pregnant females were injected with doxycycline intraperitonally at E14.5. Doxycycline then induced the transcription soluble FGFR2b in the embryos, which inhibited FGF signalling by sequestering FGF ligands, preventing them from binding to the native FGFR2b receptor. We harvested embryonic lungs after 9 hrs of inhibition. We took the left lobe of each sample for RNA isolation. We set-up our matings so that 50% of the embryos in a litter were expected to contain the soluble FGFR2b. The remaining littermates will serve as the control group. We will compare the gene expression profiles of the experimental embryonic lungs with those of littermate controls.

2020-06-22 | GSE115876 | GEO

In-vivo inhibition of Fgfr2b signalling in embryonic whole mouse lungs at E16.5

Project description:In this experiment we inhibited FGF signalling in E16.5 mouse lungs by using a dominant negative mouse model. We are interested in assessing the genetic regulation by FGF signalling at this stage of lung development. Specifically, we used a tetracycline controlled transcriptional activation model (Tet(o)) to manipulate FGF signalling. Genetically modified male mice that expressed a reverse tetracycline-controlled trans activator (rtTA) on the Rosa26 locus and a soluble FGFR2b on the Tet(o) promoter (Tet(o)sFgfr2b) (B6.Cg-Gt(ROSA)26Sortm1.1(rtTA,EGFP)NagyTg(tetO-Fgfr2b/Igh)1.3Jaw/sbel), were crossed with females lacking the Tet(o)sFgfr2b allele. Pregnant females were injected with doxycycline intraperitonally at E16.5. Doxycycline then induced the transcription soluble FGFR2b in the embryos, which inhibited FGF signalling by sequestering FGF ligands, preventing them from binding to the native FGFR2b receptor. We harvested embryonic lungs after 9 hrs of inhibition. We took the left lobe of each sample for RNA isolation. We set-up our matings so that 50% of the embryos in a litter were expected to contain the soluble FGFR2b. The remaining littermates will serve as the control group. We will compare the gene expression profiles of the experimental embryonic lungs with those of littermate controls.

2021-06-01 | GSE115877 | GEO

A rapid and scalable system for studying gene function in mice using conditional RNA interference.

Project description:RNA interference is a powerful tool for studying gene function, however, the reproducible generation of RNAi transgenic mice remains a significant limitation. By combining optimized fluorescence-coupled miR30-based shRNAs with high efficiency ES cell targeting, we developed a fast, scalable pipeline for the production of shRNA transgenic mice. Using this system, we generated eight tet-regulated shRNA transgenic lines targeting Firefly and Renilla luciferases, Oct4 and tumor suppressors p53, p16(INK4a), p19(ARF) and APC and demonstrate potent gene silencing and GFP-tracked knockdown in a broad range of tissues in vivo. Further, using an shRNA targeting APC, we illustrate how this approach can identify predicted phenotypes and also unknown functions for a well-studied gene. In addition, through regulated gene silencing we validate APC/Wnt and p19(ARF) as potential therapeutic targets in T cell acute lymphoblastic leukemia/lymphoma and lung adenocarcinoma, respectively. This system provides a cost-effective and scalable platform for the production of RNAi transgenic mice targeting any mammalian gene. PAPERCLIP:

| S-EPMC3244080 | biostudies-literature

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