scholarly journals Reporter gene assay for membrane fusion of extracellular vesicles

2021 ◽  
Vol 10 (13) ◽  
Author(s):  
Masaharu Somiya ◽  
Shun'ichi Kuroda
Keyword(s):  
Membrane Fusion ◽  
Reporter Gene ◽  
Extracellular Vesicles ◽  
Reporter Gene Assay ◽  
Gene Assay

scholarly journals Reporter gene assay for membrane fusion of extracellular vesicles

2021 ◽  
Author(s):  
Masaharu Somiya ◽  
Shun’ichi Kuroda
Keyword(s):  
Membrane Fusion ◽  
Reporter Gene ◽  
Extracellular Vesicles ◽  
Underlying Mechanism ◽  
Reporter Gene Assay ◽  
Membrane Fusion Protein ◽  
Tetracycline Transactivator ◽  
Cargo Delivery ◽  
Gene Assay ◽  
Delivery Efficiency

AbstractExtracellular vesicles (EVs) secreted by living cells are expected to deliver biological cargo molecules, including RNA and proteins, to the cytoplasm of recipient cells. There is an increasing need to understand the mechanism of intercellular cargo delivery by EVs. However, the lack of a feasible bioassay has hampered our understanding of the biological processes of EV uptake, membrane fusion, and cargo delivery to recipient cells. Here, we describe a reporter gene assay that can measure the membrane fusion efficiency of EVs during cargo delivery to recipient cells. When EVs containing tetracycline transactivator (tTA)-fused tetraspanins are internalized by recipient cells and fuse with cell membranes, the tTA domain is exposed to the cytoplasm and cleaved by protease to induce tetracycline responsive element (TRE)-mediated reporter gene expression in recipient cells. This assay (designated as EV-mediated tetraspanin-tTA delivery assay, ETTD assay), enabled us to assess the cytoplasmic cargo delivery efficiency of EVs in recipient cells. With the help of a vesicular stomatitis virus-derived membrane fusion protein, the ETTD assay could detect significant enhancement of cargo delivery efficiency of EVs. Furthermore, the ETTD assay could evaluate the effect of potential cargo delivery enhancers/inhibitors. Thus, the ETTD assay may contribute to a better understanding of the underlying mechanism of the cytoplasmic cargo delivery by EVs.

scholarly journals In vitro bioanalytical assessment of toxicity of wetland samples from Spanish Mediterranean coastline

2021 ◽  
Vol 33 (1) ◽  
Author(s):  
Alberto Celma ◽  
Geeta Mandava ◽  
Agneta Oskarsson ◽  
Juan Vicente Sancho ◽  
Lubertus Bijlsma ◽  
...  
Keyword(s):  
Water Quality ◽  
Reporter Gene ◽  
Biological Activities ◽  
Reporter Gene Assay ◽  
Water Bodies ◽  
Good Tool ◽  
Adverse Health Effects ◽  
Aryl Hydrocarbon ◽  
Gene Assay

Abstract Background Fresh water bodies represent less than 1% of overall amount of water on earth and ensuring their quality and sustainability is pivotal. Although several campaigns have been performed to monitor the occurrence of micropollutants by means of chemical analysis, this might not cover the whole set of chemicals present in the sample nor the potential toxic effects of mixtures of natural and anthropogenic chemicals. In this sense, by selecting relevant toxicity endpoints when performing in vitro bioanalysis, effect-based methodologies can be of help to perform a comprehensive assessment of water quality and reveal biological activities relevant to adverse health effects. However, no prior bioanalytical study was performed in wetland water samples from the Spanish Mediterranean coastline. Methods Eleven samples from relevant water bodies from the Spanish Mediterranean coastline were collected to monitor water quality on 8 toxicity endpoints. Aryl hydrocarbon receptor (AhR), androgenicity (AR+ and AR−), estrogenicity (ER+ and ER−), oxidative stress response (Nrf2) and vitamin D receptor (VDR+ and VDR−) reporter gene assays were evaluated. Results AhR was the reporter gene assay showing a more frequent response over the set of samples (activated by 9 out of 11 samples), with TCDD-eq in the range 7.7–22.2 pM. For AR, ER and VDR assays sporadic activations were observed. Moreover, no activity was observed on the Nrf2 reporter gene assay. Wastewater and street runaway streams from Valencia could be responsible for enhanced activities in one of the water inputs in the Natural Park ‘L’Albufera’. Conclusions Water quality of relevant wetlands from the Spanish Mediterranean coastline has been evaluated. The utilization of a panel of 5 different bioassays to cover for different toxicity endpoints has demonstrated to be a good tool to assess water quality.

Evaluation of estrogenic activities of pesticides using an in vitro reporter gene assay

2005 ◽  
Vol 15 (4) ◽  
pp. 271-280 ◽  
Author(s):  
Mihoko Kojima ◽  
Kenji Fukunaga * ◽  
Mari Sasaki ◽  
Masafumi Nakamura ◽  
Motohiro Tsuji ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Reporter Gene Assay ◽  
Gene Assay

Development of a novel reporter gene assay to evaluate antibody-dependent cellular phagocytosis for anti-CD20 therapeutic antibodies

2021 ◽  
Vol 100 ◽  
pp. 108112
Author(s):  
Chunyu Liu ◽  
Chuanfei Yu ◽  
Yalan Yang ◽  
Jing Huang ◽  
Xiaojuan Yu ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Reporter Gene Assay ◽  
Therapeutic Antibodies ◽  
Gene Assay ◽  
Anti Cd20

Dinucleotide tag‐based parallel reporter gene assay method enables efficient identification of regulatory mutations

2021 ◽  
pp. 2100341
Author(s):  
Naixia Ren ◽  
Bo Li ◽  
Qingqing Liu ◽  
Lele Yang ◽  
Xiaodan Liu ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Reporter Gene Assay ◽  
Assay Method ◽  
Gene Assay ◽  
Regulatory Mutations

Development and validation of a reporter gene assay to determine the bioactivity of anti-CTLA-4 monoclonal antibodies

2021 ◽  
Vol 101 ◽  
pp. 108277
Author(s):  
Chunyu Liu ◽  
Chuanfei Yu ◽  
Yalan Yang ◽  
Yongfei Cui ◽  
Feng Zhang ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Reporter Gene ◽  
Reporter Gene Assay ◽  
Gene Assay ◽  
Development And Validation

scholarly journals Mechanism Profiling of Hepatotoxicity Caused by Oxidative Stress Using Antioxidant Response Element Reporter Gene Assay Models and Big Data

2016 ◽  
Vol 124 (5) ◽  
pp. 634-641 ◽  
Author(s):  
Marlene Thai Kim ◽  
Ruili Huang ◽  
Alexander Sedykh ◽  
Wenyi Wang ◽  
Menghang Xia ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Big Data ◽  
Reporter Gene ◽  
Antioxidant Response ◽  
Reporter Gene Assay ◽  
Antioxidant Response Element ◽  
Response Element ◽  
Gene Assay

Validation of a reporter gene assay for the detection of synthetic glucocorticoids in calf urine samples

2014 ◽  
Vol 229 ◽  
pp. S181-S182
Author(s):  
Danilo Pitardi ◽  
Daniela Meloni ◽  
Fabio Olivo ◽  
Maria Cavarretta ◽  
Daniela Loprevite ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Reporter Gene Assay ◽  
Urine Samples ◽  
Gene Assay ◽  
Synthetic Glucocorticoids

Evaluation of Estrogenic Activity of Wastewater: Comparison Among In Vitro ERα Reporter Gene Assay, In Vivo Vitellogenin Induction, and Chemical Analysis

2015 ◽  
Vol 49 (10) ◽  
pp. 6319-6326 ◽  
Author(s):  
Masaru Ihara ◽  
Tomokazu Kitamura ◽  
Vimal Kumar ◽  
Chang-Beom Park ◽  
Mariko O. Ihara ◽  
...  
Keyword(s):  
Chemical Analysis ◽  
Reporter Gene ◽  
Estrogenic Activity ◽  
Reporter Gene Assay ◽  
Gene Assay

scholarly journals Detection of Aryl Hydrocarbon Receptor Activation by Some Chemicals in Food Using a Reporter Gene Assay

Foods ◽  
2016 ◽  
Vol 5 (1) ◽  
pp. 15 ◽  
Author(s):  
Yoshiaki Amakura ◽  
Tomoaki Tsutsumi ◽  
Morio Yoshimura ◽  
Masafumi Nakamura ◽  
Hiroshi Handa ◽  
...  
Keyword(s):  
Aryl Hydrocarbon Receptor ◽  
Reporter Gene ◽  
Receptor Activation ◽  
Reporter Gene Assay ◽  
Aryl Hydrocarbon ◽  
Gene Assay
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