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Premature termination of in vivo reprogramming leads to cancer development through altered epigenetic regulation [RRBS]

ABSTRACT: We report a reprogrammable mouse system in which reprogramming factor expression in vivo can be controlled temporally by treatment with doxycycline (Dox). Transient expression of reprogramming factors in vivo results in tumor development in various tissues, consisting of undifferentiated dysplastic cells. We analyzed the kidney tumors developed in reprogrammable mice for global gene expressions and DNA methylations.

ORGANISM(S): Mus musculus

PROVIDER: GSE52303 | GEO | 2014/02/13

SECONDARY ACCESSION(S): PRJNA227898

REPOSITORIES: GEO

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Premature termination of in vivo reprogramming leads to cancer development through altered epigenetic regulation [RRBS]

Project description:We report a reprogrammable mouse system in which reprogramming factor expression in vivo can be controlled temporally by treatment with doxycycline (Dox). Transient expression of reprogramming factors in vivo results in tumor development in various tissues, consisting of undifferentiated dysplastic cells. We analyzed the kidney tumors developed in reprogrammable mice for global gene expressions and DNA methylations. Reprogrammable mice at 4 weeks of age were treated with Dox for 7 days followed by the withdrawal. Seven days after the withdrawal, kidney tumors were analyzed for gene expressions and DNA methylations with microarray and RRBS method, respectively. Normal kidney tissue at the same age and ES cells were analyzed as controls. To examine the early changes of gene expressions, transgene-expressing kidney cells were FACS sorted and they are utilized for microarray analysis. Primary liver tumors in reprogrammable mice and transplanted secondary kidney tumors in the subcutaneous tissues of immnodeficient mice were also analyzed for gene expressions.

2014-02-13 | E-GEOD-52303 | biostudies-arrayexpress

Premature termination of in vivo reprogramming leads to cancer development through altered epigenetic regulation [array]

2014-02-13 | GSE51786 | GEO

Premature termination of in vivo reprogramming leads to cancer development through altered epigenetic regulation [array]

2014-02-13 | E-GEOD-51786 | biostudies-arrayexpress

Single-cell RNA expression profiling of mouse common lymphoid progenitors with or without dox-induction of LIN28B

Project description:To reprogram adult HSPCs in vivo, we used a mouse engineered to express LIN28B in a Doxycycline (Dox)-inducible manner (iLIN28B). We performed single cell RNA-seq to compare the transcriptomes of common lymphoid progenitor (CLP) cells isolated from mouse FL, adult BM of wildtype mice, adult BM of iLIN28B treated with Dox (+Dox), and adult BM of littermate controls +Dox.

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The role of AF10 (Mllt10) in maintaining cell identity (RNA-Seq)

Project description:AF10 is a cofactor of the H3K79 methyltransferase DOT1L. To uncover the role of H3K79me in reprogramming to induced pluripotent stem cells (iPSCs), we bred reprogrammable mice encoding doxycycline inducible Oct4-2A-Klf4-2A-IRES-Sox2-2A-c-Myc (OKSM) transgene with conditional flox (fl)-AF10 (Mllt10). We isolated mouse embryonic fibroblasts (MEFs), deleted AF10 with CRE-recombinase or treated cells with empty vector control, and initiated reprogramming in DOT1L chemical inhibitor SGC0946 or DMSO control. Gene expression was assessed using RNA-Seq and genome wide localization of H3K79me1 and H3K79me2 was determined by ChIP-Seq on day 4 of reprogramming.

2023-04-01 | GSE214139 | GEO

Effect of doxycycline treatment on germ-free mice

Project description:To characterize the mitochondrial stress response induced by Doxycycline in vivo in liver and kidneys, we administered doxycycline (Dox) at 500 mg/kg/day (mpkd) in the drinking water to 9 weeks-old germ-free C57BL/6J mice for 16 days, hence eliminating the potential confounding impacts of Doxycycline on the microbiome.

2022-09-14 | GSE202754 | GEO

Deterministic direct reprogramming of somatic cells to pluripotency [RRBS]

Project description:Somatic cells can be directly reprogrammed to pluripotency by exogenous expression of transcription factors, classically Oct4, Sox2, Klf4 and c-Myc. While distinct types of somatic cells can be reprogramed with varying efficiencies and by different modified reprogramming protocols, induced pluripotent stem cell (iPSC) induction remains inefficient and stochastic where a fraction of the cells converts into iPSCs. The nature of rate limiting barrier(s) preventing majority of cells to convert into iPSCs remains elusive. Here we show that neutralizing Mbd3, a core member of the Mbd3/NURD co-repressor and chromatin-remodeling complex, results in deterministic and synchronized reprogramming of multiple differentiated cell types to pluripotency. 100% of Mbd3 depleted mouse and human somatic cells convert into iPSCs after seven days of reprogramming factor induction. Our findings delineate a critical pathway blocking the reestablishment of pluripotency, and offer a novel platform for future dissection of epigenetic dynamics leading to iPSC formation at high resolution. Reduced representation bisulfite sequencing (RRBS) was applied to mouse iPS cells and mouse embryonic fibroblast (MEF) before and after DOX induction (initiating reprogramming by OSKM factors) from randomly selected Mbd3+/+ and Mbd3flox/- clonal cell line series. Polyclonal donor cell cultures were harvested at days 0,4 and 8 after DOX reprogramming without selection or sorting for any marker or passaging, and mapped for similarity to subcloned iPSC lines.

2014-12-13 | E-GEOD-64115 | biostudies-arrayexpress

The role of AF10 (Mllt10) in maintaining cell identity (ChIP-Seq)

Project description:AF10 is a cofactor of the H3K79 methyltransferase DOT1L. To uncover the role of H3K79me in reprogramming to induced pluripotent stem cells (iPSCs), we bred reprogrammable mice encoding doxycycline inducible Oct4-2A-Klf4-2A-IRES-Sox2-2A-c-Myc (OKSM) transgene with conditional flox (fl)-AF10 (Mllt10). We isolated mouse embryonic fibroblasts (MEFs), deleted AF10 with CRE-recombinase or treated cells with empty vector control, and initiated reprogramming in DOT1L chemical inhibitor SGC0946 or DMSO control. Gene expression was assessed using RNA-Seq and genome wide localization of H3K79me1, H3K79me2, and RNA Polymerase II (RNAPII) was determined by ChIP-Seq on day 4 of reprogramming.

2023-04-01 | GSE214141 | GEO

Microarray analysis of EpiSCs undergoing reprogramming by Esrrb.

Project description:To characterize the reprogramming of epiblast stem cells (EpiSCs) into embryonic stem cells (ESCs) induced by Esrrb, we performed microarray analysis of Tet-on Esrrb EpiSCs after treatment with doxycycline (Dox).

2018-06-01 | E-MTAB-5341 | biostudies-arrayexpress

RNA-seq analysis of doxycycline-regulated gene expression in Aspergillus fumigatus cells containing a dox off-ffmA fusion gene.

Project description:To determine the genes responsive to FfmA in A. fumigatus, a doxycycline-repressible (dox off) gene fusion to the ffmA coding sequence was constructed. This strain was designated SPF182. These cells were grown for 16 hours in the absence of doxycycline and then shifted into medium containing doxycycline (+dox) or untreated (-dox). After 8 additional hours, the cells were harvested and total RNA prepared. Wild-type cells were grown in parallel as a control in -dox medium.

2023-07-26 | GSE229912 | GEO

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