• The targeting relationships between miR-9-3p and FOXC1/FAM171A1, miR-135b-3p and RGMA were validated by dual-luciferase reporter assays. (nature.com)
  • The 3′UTR-binding sequence site of miR-128-5p on the p21 gene was predicted and assessed by luciferase assays. (hindawi.com)
  • These features make these luciferases ideal for repeated assays, sensitive measurements in miniaturized assays and single cell applications. (neb.com)
  • Luciferase assays and online miRNA databases were used to validate that tumor protein p53 (TP53) is the target gene of miR‑125b. (spandidos-publications.com)
  • To understand how the B 1 and B 2 regions influenced the transcriptional functions of nuclear HER3, we performed cyclin D1 promoter-luciferase assays in which HER3 deleted of the B 1 and B 2 regions was severely hindered in regulating this promoter. (plos.org)
  • Additional concentrated cell lysis buffer that is designed for use with Biotium's Firefly Luciferase Assay Kits when performing large sample volume assays. (biotium.com)
  • The robust signal, sensitivity, wide dynamic range, and resistance to photobleaching has made luciferase a common choice in assays ranging from use as a reporter gene in vitro and in vivo to use in biosensors. (addgene.org)
  • Luciferase reporter assays demonstrated that the Adcy1 promoter is selectively activated by neuronal PAS-domain protein 2 (NPAS2)/BMAL1. (jneurosci.org)
  • MicroRNA chip analyses, RT-PCR, gene transfection, CCK8, wound healing and Transwell assays, colony-forming assay, western blot, and the Dual-Luciferase Reporter Assay System were used to evaluate the role of miR-4778-3p in cervical cancer radiosensitivity and its relationships with target molecules NR2C2 and Med19. (bvsalud.org)
  • Background Renilla Luciferase reporter gene (rLuc) GL4. (biotech-angels.com)
  • Including a co-transfected plasmid encoding the Renilla luciferase gene ( Rluc ) in this analysis showed that its calculated stability was not as good as the optimised chromosomal genes, most likely as a result of the lower expression levels and transfection variability. (biomedcentral.com)
  • Interestingly, HLD enzymes display remarkable sequence and structural similarity with luciferase from the marine invertebrate Renilla reniformis (RLuc), which reflects their common evolutionary history. (europa.eu)
  • Unlike HLDs, which belong to the family of α/β hydrolases (EC 3.8.1.5) the RLuc luciferase is cofactor-independent monooxygenase (EC 1.13.12.5) that oxidoreductively converts substrate - coelenterazine − into coelenteramide and carbon dioxide, followed by an emission of blue light (470 nm). (europa.eu)
  • For this bioluminescent effect, the RLuc luciferase is commonly used as a reporter enzyme in cell biological research and bioimaging technologies. (europa.eu)
  • This project aimed to employ in-lab reconstructed dual-function (dehalogenase/luciferase) ancestral enzyme (ancHLD-RLuc) to decipher molecular evolution steps leading to the functional divergence of modern-day HLD and RLuc enzymes, and to explore how this knowledge could be exploited biotechnologically. (europa.eu)
  • One of the major intentions of this project was to determine atomic-level structures of the dual-function ancestral enzyme ancHLD-RLuc complexed with bound substrate molecules, with emphasis on coelenterazine-powered monooxygenation (luciferase) catalysis. (europa.eu)
  • Renilla Luciferase (RLuc), like most marine luciferases, uses a different luciferin, the benzylimidazo-pyrazinone coelenterazine. (neb.com)
  • The most common types of luciferase found in Addgene's collection are Firefly luciferase (Fluc), Renilla luciferase (Rluc), and NanoLuc® Luciferase (Nluc). (addgene.org)
  • Here we track the role of dynamics in evolution, starting from the evolvable and thermostable ancestral protein Anc(HLD-RLuc) which catalyses both dehalogenase and luciferase reactions. (muni.cz)
  • Polymerase activity was calculated by standardization of the firefly luciferase activity to the Renilla luciferase activity. (cdc.gov)
  • The ratio of Renilla and firefly luciferase activity was normalized against the unmodified plasmid psiCHECK-2-TTG. (molvis.org)
  • The relative ratio of Renilla /firefly luciferase activity determined from cells transfected with the plasmid psiCHECK-2-α was set at 1. (molvis.org)
  • The relative ratio of Renilla /firefly luciferase activity was significantly lower for the β5′-UTR than for the α5′-UTR (0.71±0.01 and 1.00±0.01, respectively). (molvis.org)
  • Three vector constructs (the unmodified plasmid psiCHECK-2-TTG and plasmids psiCHECK-2-α and -β) were transfected into HEK293 cells, and the firefly and Renilla luciferase activities were measured 24 h after transfection. (molvis.org)
  • Scientists have developed a wide range of luciferase-based plasmid systems for many different applications. (addgene.org)
  • The luciferase activities in the A549 cells incubated at 33 or 37°C and the DF-1 cells incubated at 39°C were measured by using the Dual-Glo Luciferase Assay System (Promega) at 24 h post-transfection. (cdc.gov)
  • The secreted luciferases derived from Gaussia princeps , Metridia longa and Cypridina noctiluca do not require cell lysis to measure activity and are significantly "brighter" than firefly and Renilla . (neb.com)
  • Naturally secreted from the deep sea copepod Gaussia princeps (3) (Figure 1), Gaussia Luciferase (GLuc) was first used in a sensitive analytical assay in 2002 (4) and later commercialized by NEB for expression in mammalian cells. (neb.com)
  • Other luciferases in our collection include Gaussia luciferase (Gluc) and the red and green Click-beetle luciferases (CBRluc and CBGluc, respectively). (addgene.org)
  • Gaussia luciferase is secreted and more stable, making time course experiments possible, but its short luminescence half-life makes it too dim for many experimental systems. (addgene.org)
  • The results obtained also reveal structural features distinguishing flash-type from glow-type bioluminescence, providing insights that will guide the engineering of next-generation luciferase-luciferin pairs for ultrasensitive optical bioassays. (muni.cz)
  • Luciferase is a generic term for the class of oxidative enzymes that produce bioluminescence, and is usually distinguished from a photoprotein. (wikipedia.org)
  • Luciferases are widely used in biotechnology, for bioluminescence imaging microscopy and as reporter genes, for many of the same applications as fluorescent proteins. (wikipedia.org)
  • Other state-of-the-art in vivo imaging techniques using bioluminescence imaging with luciferase-tagged pathogens and intravital 2-photon imaging systems are being evaluated for study of Chikungunya infection's chronic phase. (medscape.com)
  • Luciferase is the enzyme responsible for the bioluminescence found in a diverse number of organisms, ranging from bacteria to aquatic animals to insects. (addgene.org)
  • A modified firefly luciferase engineered to use the red-shifted substrate Akalumine with significantly brighter signal and emission in the near-infrared range, enabling enhanced bioluminescence imaging in vivo . (addgene.org)
  • Here the authors report a protein-engineering framework based on InDel mutagenesis and fragment transplantation resulting in greater catalysis and longer glow-type bioluminescence of the ancestral luciferase. (muni.cz)
  • A luciferase assay demonstrated that transfection of lens epithelial cells with wild type TP53 3'‑UTR significantly reduced the luciferase activity of the miR‑125b overexpressing cells compared with scramble controls. (spandidos-publications.com)
  • In addition, Addgene has many luciferase expression constructs available that are driven by a strong constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes in cell physiology, or as a reference level of expression. (addgene.org)
  • Here we decipher Renilla-type catalysis through crystallographic, spectroscopic and computational experiments. (muni.cz)
  • The Metridia longa secreted luciferase gene encodes a 24 kDa protein containing an N-terminal secretory signal peptide of 17 amino acid residues. (wikipedia.org)
  • Before inserting the 5′-UTR sequences into the psiCHECK-2 vector, the ATG of the Renilla luciferase was mutated to TTG, so that the Renilla luciferase expression would be driven by the primary ATG initiation codon (reverse triangle) of the gene under investigation. (molvis.org)
  • Luciferase-containing plasmids are commonly used to investigate the effect of regulatory elements, such as promoters, enhancers and untranslated regions, or the effect of mutations of these regulatory elements on gene expression. (addgene.org)
  • As targets of siRNA, the entire bPRNP coding sequence was connected to the reporter gene of the fluorescent EGFP, or of firefly luciferase or Renilla luciferase. (scientistlive.com)
  • Dinoflagellate luciferase is a multi-domain eukaryote protein, consisting of an N-terminal domain, and three catalytic domains, each of which preceded by a helical bundle domain. (wikipedia.org)
  • The structure of the dinoflagellate luciferase catalytic domain has been solved. (wikipedia.org)
  • The name was first used by Raphaël Dubois who invented the words luciferin and luciferase, for the substrate and enzyme, respectively. (wikipedia.org)
  • GLuc is a monomeric enzyme of only 185 amino acids (19 kD), and is the smallest known luciferase. (neb.com)
  • Firefly luciferase is considerably brighter than Renilla luciferase, but is more susceptible to enzyme inhibition. (addgene.org)
  • The first generation of luciferase reporters developed were based on Firefly Luciferase (FLuc). (neb.com)
  • Structures of ancestral and extant luciferases complexed with the substrate-like analogue azacoelenterazine or a reaction product were obtained, providing molecular snapshots of coelenterazine-to-coelenteramide oxidation. (muni.cz)
  • However, unlike fluorescent proteins, luciferases do not require an external light source, but do require addition of luciferin, the consumable substrate. (wikipedia.org)
  • The substrate is then available for oxidation by the luciferase, where it is degraded to coelenteramide with a resultant release of energy. (wikipedia.org)
  • There is a large pocket in the β-barrel of the dinoflagellate luciferase at pH 8 to accommodate the tetrapyrrole substrate but there is no opening to allow the substrate to enter. (wikipedia.org)
  • Since light emission depends strictly on the chemical reaction between the substrate and the luciferase, there is no background light originating from the sample. (neb.com)
  • Luciferase catalyzes the oxidation of a substrate, leading to the emission of photons. (addgene.org)
  • The ATP Cell Viability Luciferase Assay is a flash-type luminescence assay designed for individual sample detection by using a luminometer in a single sample format or a luminometer with an injector in 96-well plate format. (sigmaaldrich.com)
  • Insect luciferases, including Photinus pyralis (firefly), catalyze the oxidation of a benzothiazol luciferin requiring ATP as a cofactor. (neb.com)
  • The majority of studied luciferases have been found in animals, including fireflies, and many marine animals such as copepods, jellyfish, and the sea pansy. (wikipedia.org)
  • Renilla l uciferase was expressed under CMV promoter, A Puromycin antibiotic marker was expressed under Rsv promoter. (gentarget.com)
  • Yeast one-hybrid (Y1H) experiments suggested that LsARF3 could physically interact with the LsCO promoter, which was further confirmed by a dual luciferase assay in tobacco leaves. (frontiersin.org)
  • Pre-made lentivirus express Firefly luciferase reporter under the human CD43 promoter which expressed on the surface of leukocytes and platelets. (reportergene.com)
  • GenTargetDescription:Pre-made lentiviral particles having Renilla luciferase reporter under heat inducible Hsp70 promoter. (reportergene.com)
  • In addition, the luciferase activity of miR‑125b overexpressing cells transfected with the construct containing the rs78378222 polymorphism minor allele was also reduced compared with cells transfected with the wild type 3'‑UTR. (spandidos-publications.com)
  • Here we investigated whether a quick version of luciferase immunoprecipitation systems (QLIPS) could be used to produce a rapid, specific and quantitative test to detect autoantibodies associated with SjS. (biomedcentral.com)
  • The binding sites of miR-128-3p and TGFBR2 were predicted by Targetscan online software and confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. (degruyter.com)
  • One well-studied luciferase is that of the Photinini firefly Photinus pyralis, which has an optimum pH of 7.8. (wikipedia.org)
  • In this organism, the luciferase (Renilla-luciferin 2-monooxygenase) is closely associated with a luciferin-binding protein as well as a green fluorescent protein (GFP). (wikipedia.org)
  • The N-terminal domain is conserved between dinoflagellate luciferase and luciferin binding proteins (LBPs). (wikipedia.org)
  • The widely used coelenterazine-powered Renilla luciferase was discovered over 40 years ago, but the oxidative mechanism by which it generates blue photons remains unclear. (muni.cz)
  • One such example is the Metridia coelenterazine-dependent luciferase (MetLuc, A0A1L6CBM1) that is derived from the marine copepod Metridia longa. (wikipedia.org)
  • The catalyzed reaction is: coelenterazine + O2 → coelenteramide + CO2 + photon of light Newer luciferases have recently been identified that, unlike other luciferases, are naturally secreted molecules. (wikipedia.org)
  • Our crystallographic findings provided unprecedented molecular views of the Renilla-type luciferases complexed with a non-oxidizable coelenterazine analogue, coelenteramide and coelenteramine, revealing as-yet-unseen molecular details of bioluminescent reaction. (europa.eu)
  • NanoLuc® Luciferase is small, bright, and a secreted version of the protein is available - these attributes have made it popular with scientists since its release. (addgene.org)
  • To facilitate the identification of potential antivirals, we developed two reporter-expressing ZIKVs, each capable of expressing an enhanced green fluorescent protein or an improved luminescent NanoLuc luciferase. (mdpi.com)
  • Light emission in some bioluminescent bacteria utilizes 'antenna' such as lumazine protein to accept the energy from the primary excited state on the luciferase, resulting in an excited lulnazine chromophore which emits light that is of a shorter wavelength (more blue), while in others use a yellow fluorescent protein (YFP) with flavin mononucleotide (FMN) as the chromophore and emits light that is red-shifted relative to that from luciferase. (wikipedia.org)
  • Luciferases have become indispensable for the study of fundamental cellular processes including the regulation of transcription and translation, mRNA and protein stability, and nucleic acid/protein interactions. (neb.com)
  • Medium-Throughput Detection of Hsp90/Cdc37 Protein-Protein Interaction Inhibitors Using a Split Renilla Luciferase-Based Assay. (nih.gov)
  • Both Firefly luciferase and Renilla luciferase have short protein half-lives, making them useful as transcriptional reporters. (addgene.org)
  • Firefly and Renilla luciferase are less-than-ideal protein tags due to their large size. (addgene.org)
  • We evaluated FOXP1 activity by an in vitro luciferase model and assessed protein stability in vitro by western blotting. (bmj.com)
  • As for their wild-type counterparts, fusion proteins linking CRLR and RAMP1 to the energy donor Renilla luciferase ( R luc) and energy acceptor green fluorescent protein (GFP) reach the cell surface only upon coexpression of CRLR and RAMP1. (acs.org)
  • However, luciferases have been studied in luminous fungi, like the Jack-O-Lantern mushroom, as well as examples in other kingdoms including bioluminescent bacteria, and dinoflagellates. (wikipedia.org)
  • Recently, a new generation of bioluminescent reporters with unique characteristics that expand the flexibility and applications of luciferase reporter systems has become available. (neb.com)
  • Although both of these intracellular luciferases are widely used in life science research, they have the disadvantage of requiring lysis of the expressing cells in order to measure their activity. (neb.com)
  • As expected clones varied in their specific luciferase activity along with the dilutions. (biotech-angels.com)
  • Luciferase activity. (molvis.org)
  • Luc2 is the second-generation version of Firefly luciferase that has been codon optimized for expression in mammalian systems. (addgene.org)
  • Like Firefly luciferase, Click-beetle red luciferase has also been codon optimized ( CBR2opt ) for mammalian expression systems. (addgene.org)
  • A minigenome assay based on the dual-luciferase system was performed as previously reported ( 2 , 3 ). (cdc.gov)
  • Next, nine HER3-CTD truncation mutants were constructed to map minimal regions of transactivation potential using the Gal4 UAS-luciferase based system. (plos.org)
  • A variety of organisms regulate their light production using different luciferases in a variety of light-emitting reactions. (wikipedia.org)
  • An anisotropic network model highlights the importance of the conformational flexibility of a loop-helix fragment of Renilla luciferases for ligand binding. (muni.cz)
  • Empty backbones for inserting regulatory elements before or after the luciferase ORF can be found in a table below, as well as plasmids that already contain a region of interest before or after luciferase . (addgene.org)
  • Furthermore, a Dual‑Luciferase reporter assay indicated that SRY‑box 4 (SOX4) is a direct target of miR‑320, and the restoration of SOX4 in miR‑320‑overexpressing cells attenuated the tumor‑suppressive effects of miR‑320. (spandidos-publications.com)
  • He worked at Promega Corp. developing novel substrates for live-cell applications using Renilla luciferase. (hstalks.com)
  • This light emission can be easily detected and measured, making luciferases ideal for the development of biological reporter systems. (neb.com)
  • A set of three sensors that are increasingly structurally destabilized versions of Firefly luciferase which serves as a convenient toolkit to assess the proteostasis status in a wide range of experimental systems. (addgene.org)
  • ATP Cell Viability Luciferase Assay A highly sensitive firefly luciferase cell based assay for quantifying ATP in cell cultures used to measure cell viability. (sigmaaldrich.com)
  • The ATP Cell Viability Luciferase Assay offers a highly sensitive homogenous assay for quantifying ATP in cell cultures. (sigmaaldrich.com)
  • Store ATP Cell Viability Luciferase Assay at -80°C. Avoid repeated freeze-thaw cycles. (sigmaaldrich.com)
  • However, due to the closely associated GFP, the energy released by the luciferase is instead coupled through resonance energy transfer to the fluorophore of the GFP, and is subsequently released as a photon of green light (peak emission wavelength 510 nm). (wikipedia.org)