Thematische Bibliographien / NanoLuciferase

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Auswahl der wissenschaftlichen Literatur zum Thema „NanoLuciferase“

Autor: Grafiati

Veröffentlicht am 28. Juni 2021

Zuletzt aktualisiert am 7. Februar 2022

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Zeitschriftenartikel zum Thema "NanoLuciferase"

Sfarcic, Ivana, Theresa Bui, Erin C. Daniels und Emily R. Troemel. „Nanoluciferase-Based Method for Detecting Gene Expression in Caenorhabditis elegans“. Genetics 213, Nr. 4 (04.10.2019): 1197–207. http://dx.doi.org/10.1534/genetics.119.302655.

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Genetic reporters such as the green fluorescent protein (GFP) can facilitate measurement of promoter activity and gene expression. However, animal autofluorescence limits the sensitivity of GFP and other fluorescent reporters in whole-animal settings like in the nematode Caenorhabditis elegans. Here, we present a highly sensitive Nanoluciferase (NanoLuc)-based method in a multiwell format to detect constitutive and inducible gene expression in C. elegans. We optimize detection of bioluminescent signals from NanoLuc in C. elegans and show that it can be detected at 400,000-fold over background in a population of 100 animals expressing intestinal NanoLuc driven by the vha-6 promoter. We can reliably detect signal in single vha-6p::Nanoluc-expressing worms from all developmental stages. Furthermore, we can detect signal from a 1/100 dilution of lysate from a single vha-6p::Nanoluc-expressing adult and from a single vha-6p::Nanoluc-expressing adult “hidden” in a pool of 5000 N2 wild-type animals. We also optimize various steps of this protocol, which involves a lysis step that can be performed in minutes. As a proof-of-concept, we used NanoLuc to monitor the promoter activity of the pals-5 stress/immune reporter and were able to measure 300- and 50-fold increased NanoLuc activity after proteasome blockade and infection with microsporidia, respectively. Altogether, these results indicate that NanoLuc provides a highly sensitive genetic reporter for rapidly monitoring whole-animal gene expression in C. elegans.

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Pulkina, A. A., E. A. Romanovskaya-Romanko, A. S. Mustafaeva, A. Yu Egorov und M. A. Stukova. „Rapid Neutralizing Antibody Assessment Using Influenza Viruses with Luciferase Reporter“. Biotekhnologiya 37, Nr. 2 (2021): 81–91. http://dx.doi.org/10.21519/0234-2758-2021-37-2-81-91.

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Influenza viruses cause an acute respiratory infection, especially in the autumn-winter period. They are characterized by a high mutation frequency and cause annual seasonal epidemics. The detection of virus-neutralizing antibodies is an important criterion in the assessment of population immunity and the influenza vaccine effectiveness. In this study, a method for determining the titer of virus-neutralizing antibodies in blood serum has been developed. A new test, called the luciferase neutralization assay, uses measurement of a bioluminescent signal as a detection method. The influenza A reporter viruses of various subtypes were constructed that encode the nanoluciferase protein in the non-structural NS1 protein reading frame. The developed method was used to compare paired sera of volunteers before and after their immunization with a seasonal influenza vaccine. The proposed method was also compared with certified antibody assay methods: neutralization reaction and hemagglutination inhibition reaction. The tests showed a high correlation, while the luciferase neutralization assay reduced the time and simplified the detection procedure. microneutralization, reporter virus, influenza virus, bioluminescence, nanoluciferase The study was supported by the grant of the President of the Russian Federation for young PhDs (no. 075-15-2019-226); and also by the grant of the Government of Saint-Petersburg for undergraduate and graduate students (September 25, 2018, no 124).

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Sahihi, Mehdi, Juan Sanz García und Isabelle Navizet. „Bioluminescent Nanoluciferase–Furimamide Complex: A Theoretical Study on Different Protonation States“. Journal of Physical Chemistry B 124, Nr. 13 (10.03.2020): 2539–48. http://dx.doi.org/10.1021/acs.jpcb.9b11597.

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Wires, Emily S., Doug Howard, Mark J. Henderson, Xiaokang Yan, Kathleen A. Trychta, Emily J. Heathward, Yajun Zhang, Molly Lutrey, Christopher Richie und Brandon K. Harvey. „218. Monitoring ER Stress Activation of the ATF6 Pathway Using Nanoluciferase“. Molecular Therapy 24 (Mai 2016): S85. http://dx.doi.org/10.1016/s1525-0016(16)33027-1.

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Degrelle, Séverine A., Hussein Shoaito und Thierry Fournier. „New Transcriptional Reporters to Quantify and Monitor PPARγ Activity“. PPAR Research 2017 (2017): 1–7. http://dx.doi.org/10.1155/2017/6139107.

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The peroxisome-proliferator-activated-receptor-γ (PPARγ) is a member of the nuclear receptor superfamily that plays a critical role in diverse biological processes, including adipogenesis, lipid metabolism, and placental development. To study the activity of PPARγ, we constructed two new reporter genes: a fluorescent GFP-tagged histone-2B (PPRE-H2B-eGFP) and a secreted nanoluciferase (PPRE-pNL1.3[secNluc]). This study demonstrates their usage to monitor PPARγ activity in different cell types and screen for PPARγ’s potential ligands.

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Calverley, Ben C., Karl E. Kadler und Adam Pickard. „Dynamic High-Sensitivity Quantitation of Procollagen-I by Endogenous CRISPR-Cas9 NanoLuciferase Tagging“. Cells 9, Nr. 9 (10.09.2020): 2070. http://dx.doi.org/10.3390/cells9092070.

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The ability to quantitate a protein of interest temporally and spatially at subcellular resolution in living cells would generate new opportunities for research and drug discovery, but remains a major technical challenge. Here, we describe dynamic, high-sensitivity protein quantitation technique using NanoLuciferase (NLuc) tagging, which is effective across microscopy and multiwell platforms. Using collagen as a test protein, the CRISPR-Cas9-mediated introduction of nluc (encoding NLuc) into the Col1a2 locus enabled the simplification and miniaturisation of procollagen-I (PC-I) quantitation. Collagen was chosen because of the clinical interest in its dysregulation in cardiovascular and musculoskeletal disorders, and in fibrosis, which is a confounding factor in 45% of deaths, including those brought about by cancer. Collagen is also the cargo protein of choice for studying protein secretion because of its unusual shape and size. However, the use of overexpression promoters (which drowns out endogenous regulatory mechanisms) is often needed to achieve good signal/noise ratios in fluorescence microscopy of tagged collagen. We show that endogenous knock-in of NLuc, combined with its high brightness, negates the need to use exogenous promoters, preserves the circadian regulation of collagen synthesis and the responsiveness to TGF-β, and enables time-lapse microscopy of intracellular transport compartments containing procollagen cargo. In conclusion, we demonstrate the utility of CRISPR-Cas9-mediated endogenous NLuc tagging to robustly quantitate extracellular, intracellular, and subcellular protein levels and localisation.

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Abe, Taisho, Riku Nagai, Hiroaki Imataka und Nono Takeuchi-Tomita. „Reconstitution of yeast translation elongation and termination in vitro utilizing CrPV IRES-containing mRNA“. Journal of Biochemistry 167, Nr. 5 (16.03.2020): 441–50. http://dx.doi.org/10.1093/jb/mvaa021.

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Abstract We developed an in vitro translation system from yeast, reconstituted with purified translation elongation and termination factors and programmed by CrPV IGR IRES-containing mRNA, which functions in the absence of initiation factors. The system is capable of synthesizing the active reporter protein, nanoLuciferase, with a molecular weight of 19 kDa. The protein synthesis by the system is appropriately regulated by controlling its composition, including translation factors, amino acids and antibiotics. We found that a high eEF1A concentration relative to the ribosome concentration is critically required for efficient IRES-mediated translation initiation, to ensure its dominance over IRES-independent random internal translation initiation.

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Kim, Jiho, und Regis Grailhe. „Nanoluciferase signal brightness using furimazine substrates opens bioluminescence resonance energy transfer to widefield microscopy“. Cytometry Part A 89, Nr. 8 (03.05.2016): 742–46. http://dx.doi.org/10.1002/cyto.a.22870.

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Zhang, Lei, Ge Song, Ting Xu, Qing-Ping Wu, Xiao-Xia Shao, Ya-Li Liu, Zeng-Guang Xu und Zhan-Yun Guo. „A novel ultrasensitive bioluminescent receptor-binding assay of INSL3 through chemical conjugation with nanoluciferase“. Biochimie 95, Nr. 12 (Dezember 2013): 2454–59. http://dx.doi.org/10.1016/j.biochi.2013.09.008.

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Ji, Ben-Jun, Ge Song, Zhou Zhang und Zhan-Yun Guo. „Efficient overexpression of human interleukin-6 in Escherichia coli using nanoluciferase as a fusion partner“. Process Biochemistry 50, Nr. 10 (Oktober 2015): 1618–22. http://dx.doi.org/10.1016/j.procbio.2015.06.008.

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Dissertationen zum Thema "NanoLuciferase"

Šlaufová, Marta. „Vzájemné interakce mezi nádorovým mikroprostředím a kalikreinovými proteázami v myším modelu karcinomu mléčné žlázy“. Master's thesis, 2021. http://www.nusl.cz/ntk/nusl-446587.

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Breast cancer is the most common cancer type with a high annual death rate. Finding meaningful tissue-related or body-fluid-accessible biomarkers is necessary to characterize cancer subtype, predict tumor behavior, choose the most effective therapy, predict severe treatment-related toxicities, and also the opportunity to personalize treatments for each patient. There is increasing evidence that various kallikrein-related peptidases (Klk) gene family members can modulate the immune response and are differentially regulated in breast cancer, and therefore are proposed to be potential prognostic biomarkers. This work established and validated an experimental setup to study the roles of selected kallikrein-related peptidases (KLK5, KLK7, KLK14) in breast cancer in vivo using gene-deficient mouse models previously generated in our laboratory. We used the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats) editing system to generate several E0771 cell line-based reporter and gene-deficient cell lines. These allowed enhanced monitoring of cancer progression in vivo and studying KLKs roles in tumor immune microenvironment of C57Bl/6N mice. Finally, we present the analysis of the initial in vivo experiments using these tools combined with established Klk-deficient mouse models. Our...

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Buchteile zum Thema "NanoLuciferase"

Laschet, Céline, und Julien Hanson. „Correction to: Nanoluciferase-Based Complementation Assay to Detect GPCR-G Protein Interaction“. In Methods in Molecular Biology, C1—C2. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1221-7_21.

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Schihada, Hannes, Katarina Nemec, Martin J. Lohse und Isabella Maiellaro. „Bioluminescence in G Protein-Coupled Receptors Drug Screening Using Nanoluciferase and Halo-Tag Technology“. In Methods in Molecular Biology, 137–47. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1221-7_9.

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Zhang, Wei, Terunao Takahara, Takuya Achiha, Hideki Shibata und Masatoshi Maki. „Cellular Ca2+-Responding Nanoluciferase Reporter Gene System Directed by Tandemly Repeated Pseudo-palindromic NFAT-Response Elements“. In Methods in Molecular Biology, 95–109. New York, NY: Springer New York, 2019. http://dx.doi.org/10.1007/978-1-4939-9030-6_7.

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Zhang, Wei, Rina Matsuo, Terunao Takahara, Hideki Shibata und Masatoshi Maki. „High Sensitive Quantitative Binding Assays Using a Nanoluciferase-Fused Probe for Analysis of ALG-2-Interacting Proteins“. In Methods in Molecular Biology, 501–16. New York, NY: Springer New York, 2019. http://dx.doi.org/10.1007/978-1-4939-9030-6_31.

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