Silencing of fat-1 transgene expression in sheep may result from hypermethylation of its driven cytomegalovirus (CMV) promoter

Biao, Duan; Liu, Cheng; Gao, Yu; Yin, Fufen; Su, Guanghua; Shen, Qi; Liu, K.; Hu, Xiao; Liu, X.; Li, G.P.

doi:10.1016/j.theriogenology.2012.03.027

Cited by 33 publications

(32 citation statements)

References 35 publications

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“…The CAG reporter uses the same enhancer as CMV, but uses a chicken β-actin promoter rather than the CMV promoter. CAG’s β-actin promoter produces more reliable expression across cell lines than the CMV promoter [21,22]. Further experiments will establish the methylation-dependency of these reporters in our culture conditions.…”

Section: Discussionmentioning

confidence: 99%

A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter

Carrico

Malinow

2017

J Biol Methods

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Robust assays for detecting the effects of elevated concentrations of amyloid-β (Aβ) may facilitate Alzheimer’s disease research. An appropriate assay would be high-throughput and enable identification of drugs and genetic mutations that block the effects of Aβ, potentially leading to treatments for Alzheimer’s disease. We discovered that the commonly used cytomegalovirus (CMV) enhancer/promoter is sensitive to the effects of Aβ. By combining the CMV enhancer/promoter with a fluorescent protein, we created a reporter system that produces changes in intracellular fluorescence in response to Aβ. Using hippocampal neurons, we quantified the ability of a CMV-fluorescent protein recombinant reporter to detect both exogenously applied and overexpressed Aβ. This is the first report of a high-throughput enhancer/promoter-based Aβ detection method. The reporter is able to detect the effects of elevated concentrations of Aβ in a high-throughput fashion, providing a new tool for Alzheimer’s disease research and important knowledge about the commonly used CMV enhancer/promoter.

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Section: Discussionmentioning

confidence: 99%

A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter

Carrico

Malinow

2017

J Biol Methods

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“…In addition, some studies had shown that promoters may be influenced by epigenetic modifications which can result in silencing of transgenes in genetically engineered animals. Recently, Duan et al (2012) reported that hypermethylated cytomegalovirus (CMV) promoter resulted in inhibition of fat-1 transgenes expression in transgenic sheep.…”

Section: Discussionmentioning

confidence: 99%

Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference

Qiao

et al. 2015

Journal of Virological Methods

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“…It is not clear from our studies why this occurred, but the main reason may be the choice of promoter in our vector system. We used the strong CMV promoter, which is able to produce enormous amounts of transgene products but has a propensity to be silenced over time (Prosch et al, 1996;Mehta et al, 2009;Duan et al, 2012). Thus, some small molecules such as DNA methyltransferase inhibitors or histone deacetylase inhibitors might be able to sustain hFIX gene expression.…”

Section: Discussionmentioning

confidence: 99%

Genetically Modified Adipose Tissue-Derived Stem/Stromal Cells, Using Simian Immunodeficiency Virus-Based Lentiviral Vectors, in the Treatment of Hemophilia B

et al. 2013

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Hemophilia is an X-linked bleeding disorder, and patients with hemophilia are deficient in a biologically active coagulation factor. This study was designed to combine the efficiency of lentiviral vector transduction techniques with murine adipose tissue-derived stem/stromal cells (mADSCs) as a new method to produce secreted human coagulation factor IX (hFIX) and to treat hemophilia B. mADSCs were transduced with simian immunodeficiency virus (SIV)-hFIX lentiviral vector at multiplicities of infection (MOIs) from 1 to 60, and the most effective dose was at an MOI of 10, as determined by hFIX production. hFIX protein secretion persisted over the 28-day experimental period. Cell sheets composed of lentiviral vector-transduced mADSCs were engineered to further enhance the usefulness of these cells for future therapeutic applications in transplantation modalities. These experiments demonstrated that genetically transduced ADSCs may become a valuable cell source for establishing cell-based gene therapies for plasma protein deficiencies, such as hemophilia.

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Silencing of fat-1 transgene expression in sheep may result from hypermethylation of its driven cytomegalovirus (CMV) promoter

Cited by 33 publications

References 35 publications

A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter

A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter

Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference

Genetically Modified Adipose Tissue-Derived Stem/Stromal Cells, Using Simian Immunodeficiency Virus-Based Lentiviral Vectors, in the Treatment of Hemophilia B

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