• We examined the activities of cloned promoter DNA fragments with luciferase assay. (molvis.org)
  • A luciferase assay using a plasmid containing the PXR response elements of CYP3A4 revealed that the increased CYP3A4 induction by siNEAT1_2 or siDAZAP1 was due to the increased transactivation by PXR. (aspetjournals.org)
  • Luciferase assay series of TRAIL promoters for the cells co-transfected with RUNX1 and core-binding factor ß (CBFß) expression vectors were performed to evaluate the nature of TRAIL transcriptional regulation. (bvsalud.org)
  • Then, we measured the promoter activity of various 5′ flanking regions of cNANOG in chicken PGCs and ESCs using the luciferase reporter assay. (biomedcentral.com)
  • A luciferase reporter assay, in A549 cells, identified allele-dependent mRNA transcription. (cdc.gov)
  • The transcriptional activity of the fragment was demonstrated in mouse Neuro-2a cells by a luciferase assay. (elsevierpure.com)
  • To determine which transcription factors bind to the promoter, we carried out a systematic search for the binding proteins using the 1142 bp Gal-T-II promoter fragment containing both positive and negative regulatory regions in a combination of DNA pull-down assay and transcription factor array analysis. (elsevierpure.com)
  • Standard techniques like cell synchronization by serum starvation, flow cytometry, N-terminal or C-terminal FLAG epitope-tagged protein expression, immunofluorescence confocal microscopy, dual luciferase assay for promoter evaluation, and chromatin immunoprecipitation assay were employed during this study. (biomedcentral.com)
  • A luciferase reporter assay, RNA immunoprecipitation (RIP), RNA pulldown assays and fluorescence in situ hybridization were performed to identify the interactions among circIGF2BP3, miR-328-3p, miR-3173-5p and plakophilin 3 (PKP3). (biomedcentral.com)
  • Approximately 8700 base pairs (bp) of the 5′-flanking region of the rat oatp2 gene were linked to the luciferase reporter gene and used in transient transfection assays in H4IIE cells. (aspetjournals.org)
  • On the basis of gel mobility shift and supershift assays, we demonstrated that both YY1 and USF1 transcription factors can bind to the 52-base pair region. (elsevierpure.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)
  • Microarrays followed by bioinformatic analyses, luciferase reporter assays and Western blotting were applied for mechanistic studies. (thno.org)
  • These miRNAs which targeted β -catenin mRNA were confirmed by dual-luciferase reporter system and RNA-pulldown. (hindawi.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)
  • To investigate the events involved in regulation of the secretogranin II (SgII) gene, luciferase reporter constructs were transfected into gonadotrope-derived, αT3-1 cells. (elsevierpure.com)
  • In addition, exogenous haem (20 microM) repressed the basal and Glut- and DHA-induced activity of luciferase reporter constructs containing 9.1 and 6.3 kb of ALAS-1 5'-flanking region but not the construct containing the first 3.5 kb of promoter sequence. (umassmed.edu)
  • 2.8 kb haem-repressive region (1.7 and 1.1 kb constructs) retained haem-dependent repression of basal and drug inductions, suggesting that more than one cis-acting elements are responsible for this haem-dependent repression of ALAS-1. (umassmed.edu)
  • Luciferase reporter constructs containing either allele were transiently transfected into HeLa cells. (ucl.ac.uk)
  • The proximal 550 bp fragment showed the highest transcriptional activity as determined by a series of 5′-truncated constructs of the promoter. (elsevierpure.com)
  • Analysis of 5' promoter deletion mutants revealed that the region from -441 to +118, relative to the transcription initiation site, is sufficient to establish high level promoter activity. (nih.gov)
  • Transient transfection analysis with deleted mutants of a luciferase reporter construct showed that the promoter activity was regulated negatively and positively in the distal and proximal promoter regions, respectively. (eurekamag.com)
  • With a series of deletion/point mutants, we identified CaM antagonist-responsive region in the putative Sp1-binding domain between -295 to -300 base pairs of DR5 gene. (oncotarget.com)
  • 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)
  • This paper reports the isolation of two putative D 2 R promoters from grey mullet, one 5′ flanking and the other an intronic sequence immediately upstream of the first coding exon. (huji.ac.il)
  • DNA methylation occurs mainly at CpG-rich 'CpG islands' and surrounding 'CpG-shore' regions, where more than 60% of them are located in 5′ promoters [ 2 ]. (biomedcentral.com)
  • To investigate the molecular mechanisms regulating sst2 gene expression, we cloned the upstream region (9.5 kb) from the translation initiation codon of the rat sst2 gene. (eurekamag.com)
  • Transient expression of a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the 52-base pair region from - 326 to -275 upstream of the transcription initiation site includes a positive cisacting element(s) for the efficient expression of hTOP3 gene. (elsevierpure.com)
  • CONCLUSION: These results indicate there are two effective gene regions that regulate hBD-2 expression by IL-1a between 2.7kbp and 1.1kbp, and 500bp upstream of the translation starting point of hBD-2 in HMEEC-1. (ajou.ac.kr)
  • It has been proposed that ribosome scanning and start codon selection are regulated by elements in the 5' leader sequence, such as RNA primary sequences (for example, the Kozak sequence context), upstream open reading frames (uORFs), secondary structures, and RNA modifications 4 - 7 . (biorxiv.org)
  • When this 400-bp region was placed upstream of a luciferase reporter (−0.4m Gdf9 - Luc ), oocyte-specific expression was observed. (bioone.org)
  • A complete HERV contains the gag, pro, pol and env genes, flanked either side by the long terminal repeats. (wikipedia.org)
  • Gene transcription is mainly regulated by transcription factors (TFs) that bind to specific DNA sequences (called motifs) located in the promoter regions of genes [ 1 ]. (biomedcentral.com)
  • DEAF-1 activates the expression of Mtk and Drs promoter-luciferase fusion genes in S2 cells. (sdbonline.org)
  • To identify regulatory elements that confer high-level expression of GDF9 in the ovary but repression in other tissues, we generated transgenic mice in which regions of the Gdf9 locus were fused to reporter genes. (bioone.org)
  • circRNAs are generated by back-splicing from their host genes and are highly stable due to the absence of 5′ caps and 3′ tails [ 11 ]. (biomedcentral.com)
  • To delimit the control elements that mediate tissue-specific and sex hormone-responsive apo[a] transcription, we derived a reporter YAC in which 40 kb of 5' flanking sequences from the cloned apo[a] allele were linked to a luciferase reporter gene. (zhihuiya.com)
  • The mechanisms involved in the stress-induced translation have been investigated for a small number of key transcription factors (for example, yeast general control nondepressible 4 (GCN4) 12 and mammalian activating transcription factor 4 (ATF4) 13 ), whose translation is normally inhibited by the uORFs in the 5' leader sequences of their mRNAs. (biorxiv.org)
  • Using in planta nucleotide-resolution mRNA structurome probing, we discovered that this stress-induced switch in translation is mediated by highly structured regions detected downstream of uAUGs in TE-up transcripts. (biorxiv.org)
  • The preinitiation complex then scans along the 5' leader sequence of the mRNA to initiate translation at a start codon 1 - 3 . (biorxiv.org)
  • The proximal region of the cNANOG promoter contains a positive transcriptional regulatory element (CCAAT/enhancer-binding protein ( CEBP )-binding site) in ESCs. (biomedcentral.com)
  • The binding sites of T-box transcription factor TBX5 and NK-2 transcription factor-related locus 5 Nkx2.5, which were expressed at higher levels in aortic smooth muscle cells than in T cells, were located within the −387 to +97 NFAT3 promoter region, exhibiting the maximum enhancer activity. (aai.org)
  • The insertion of this motif into the Diptericin ( Dpt ) regulatory region confers DEAF-1 responsiveness to this normally DEAF-1-independent enhancer. (sdbonline.org)
  • The luciferase-expressing vector containing diverse lengths of the hBD-2 5' flanking region made by the progressive unidirectional deletion was transferred to HMEEC. (ajou.ac.kr)
  • 단방향 유전자 절단을 이용하여(unidirectional DNA deletion) vector construct를 만든 뒤, 전기영동과 염기서열분석을 통해 5'flanking 부위의 크기를 알아내었고 필요한 vector construct를 충분히 확보하였으며, 각각을 transfection 시킨 뒤, luminometer를 이용, luciferase의 활성도를 측정하여 5'flanking 부위의 유전자 조절 기능을 분석하였다. (ajou.ac.kr)
  • Scientists have developed a wide range of luciferase-based plasmid systems for many different applications. (addgene.org)
  • When HeLa cells were transiently transfected with a mutant construct which had disabled both YY1- and USF1-binding sites, the luciferase activity was greatly reduced, suggesting that these binding elements play a functional role in the basal activation of the hTOP3 promoter. (elsevierpure.com)
  • A 9.1 kb cALAS-1 (chicken ALAS-1) promoter-luciferase-reporter construct, was poorly induced by Glut and not by DHA alone, but was synergistically induced by the combination. (umassmed.edu)
  • 0.005) basal expression of luciferase, compared to the G(A8T12) construct. (ucl.ac.uk)
  • Following stimulation with lipopolysaccharide or interleukin-1, luciferase expression from the C(A8T12) construct did not change. (ucl.ac.uk)
  • Rasd1 inhibited transcriptional repression by Ear2 on a renin promoter-luciferase reporter construct both in the presence and absence of all-trans-retinoic acid. (biomedcentral.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)
  • 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)
  • We also demonstrated that the −657 to +411 DCT promoter fragment efficiently directs RB cell-specific transcription of the luciferase reporter gene in cell lines. (molvis.org)
  • Single-strand conformational polymorphism analysis of a PCR-generated fragment from the 5' flanking region of the IL-6 gene (-550 to +61) indicated the existence of polymorphisms. (ucl.ac.uk)
  • To understand the transcriptional regulation of Gal-T-II gene expression, we cloned a 1448 bp 5′-flanking fragment from the mouse Gal-T-II gene. (elsevierpure.com)
  • This 5' flanking region is a functional promoter of the PDGF alpha R gene as concluded from its capacity to drive luciferase reporter gene expression in an orientation dependent way. (nih.gov)
  • These studies offer evidence that GnRH-induced activation of the SgII promoter in the αT3-1 cell line requires cAMP-dependent protein kinase activity and a functional CRE within the 5′-flanking region of the gene. (elsevierpure.com)
  • Promoter activity of the intronic sequence was confirmed in vitro through functional analysis using luciferase as reporter gene. (huji.ac.il)
  • The functional characteristics of the region flanking the 5′-UTR is currently under investigation. (huji.ac.il)
  • p ≤ 10-3) (range 10-3.006- 10-5.504, nearly all located in intronic regions. (cdc.gov)
  • The 5-year survival rate has only improved from 2% to 6% in the past 30 years [ 2 ]. (oncotarget.com)
  • Unfortunately, even among those patients who undergo resection for pancreatic cancer and have tumor-free margins, the 5-year survival rate after resection is 10% to 25% [ 3 ]. (oncotarget.com)
  • Dysregulation of apoptosis of tumor cells plays an important role in the pathogenesis of pancreatic cancer and their resistance to therapies [ 5 ]. (oncotarget.com)
  • Noncoding RNAs, including circRNAs, microRNAs, and lncRNAs, have become a significant focus of research attention due to their crucial role in regulating the progression of multiple malignancies [ 5 ], including bladder cancer, gastric cancer, nasopharyngeal carcinoma, and pancreatic cancer. (hindawi.com)
  • Methods For subcutaneous studies, 1 × 10 6 Renca cells were implanted subcutaneously (s.c.) into the right hind flank and mice were subsequently vaccinated i.p. on day 5, 12, and 19 with Lm-based vaccines. (bmj.com)
  • Briefly, 1×106 Renca cells in 100 µl PBS were implanted subcutaneously into the right hind flank and mice were subsequently vaccinated i.p. with Lm-based vaccines. (bmj.com)
  • Nude mice were subcutaneously or intramuscularly implanted with luciferase-expressing murine 1029H or human KRIB osteosarcoma cells. (biomedcentral.com)
  • To understand the mechanisms responsible for regulating hTOP3 expression, we have cloned the 5'-flanking region of the gene coding for the hTOP3 and analyzed its promoter activity. (elsevierpure.com)
  • We analyzed the function of 5' flanking region by luciferase activity measured using luminometer after corresponding substrate was supplemented to the cell lysate. (ajou.ac.kr)
  • The treatment of IL-1a upregulates the activity of promoter 7.60±1.45 (average ± standard deviation) folded in 2.7kpb sized 5' flanking region, 3.81 ± 0.78 folded in 1.1kbp, and 4.00 ± 0.73 folded in 500bp, respectively. (ajou.ac.kr)
  • A study employing NFAT1/NFAT3 chimeric molecules revealed that the region in NFAT3 responsible for NFAT promoter activity inhibition was located within its N-terminal transactivation domain, Ca 2+ -regulatory domain, and DNA-binding domain. (aai.org)
  • When the cells are in the dividing state (S/G 2 ), the forward activity of the promoter is 5-8 folds higher than the reverse activity. (biomedcentral.com)
  • We evaluated FOXP1 activity by an in vitro luciferase model and assessed protein stability in vitro by western blotting. (bmj.com)
  • The expression of TRAIL, its receptor Death receptor 4 and 5 and RUNX1 in K562 cells transfected with the RUNX1 expression vector and RUNX1 siRNA were evaluated by reverse transcription-quantitative PCR (RT-qPCR). (bvsalud.org)
  • In contrast, transgenes −10.7m Gdf9 - Luc and −3.3m Gdf9 - Luc , which lacked the downstream 5.6-kb region, demonstrated reporter expression not only in oocytes but also high expression in male germ cells. (bioone.org)
  • This suggests that the downstream 5.6-kb sequence contains a testis-specific repressor element and that 3.3 kb of 5′-flanking sequence contains all the cis -acting elements for directing high expression of Gdf9 to female (and male) germ cells. (bioone.org)
  • Flanking the CZ is the peripheral zone (PZ) where cells are recruited for the initiation of new lateral primordia. (nature.com)
  • For example, miR-552 promotes cell proliferative rate of laryngocarcinoma cells by modulating p53 signaling [ 5 ]. (hindawi.com)
  • For orthotopic studies, 5 × 10 4 Renca cells expressing luciferase (Renca-luc) were implanted directly into the right kidney as previously described and mice were subsequently vaccinated i.p. on day 5, 12, and 17 with Lm-based vaccines. (bmj.com)
  • Briefly, 5×104 Renca cells expressing luciferase (Renca-luc) were implanted directly into the left kidney and mice were subsequently vaccinated i.p. with Lm-based vaccines. (bmj.com)
  • 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)
  • DEAF-1 recognizes several TTCG motifs within the portion of the Dfd autoregulatory region termed 'module E.' In addition, DEAF-1 binds several similar motifs within a Dfd response element ( DRE ) from the 1.28 gene that enhances maxillary gene expression during embryogenesis (Pederson, 2000). (sdbonline.org)
  • A distinct ZmFCP1-FEA3 ligand-receptor combination, originally identified in maize, prevents WUS gene expression in the region below the OC (also called rib zone, RZ), thus confining WUS expression within the OC of meristems 6 . (nature.com)
  • This study describes the molecular cloning of the non-coding exon 1 and approximately 2 kb of 5' flanking region of the human PDGF alpha R gene. (nih.gov)
  • miRNAs induce gene degradation and suppression of translation, or both, mainly by imperfect binding to miRNA recognition elements (MREs) within the 3′-untranslated region (UTR) of target mRNAs ( 1 ). (frontiersin.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)
  • An expression vector for an inhibitor of the cAMP-dependent protein kinase was found to reduce the ability of cAMP or GnRH to activate the SgII-luciferase reporter gene. (elsevierpure.com)
  • Direct binding of ZFP143 to the proximal region of the NANOG promoter regulates NANOG expression by modulating OCT4 binding [ 24 ]. (biomedcentral.com)
  • These results demonstrate that there are regulatory regions in the 5'-flanking region of the cALAS-1 gene that respond to haem and provide a basis for further investigations of the molecular mechanisms by which haem down-regulates expression of the ALAS-1 gene. (umassmed.edu)
  • This thesis discusses research conducted to examine the hypothesis that this is due to polymorphisms in the region of the IL-6 gene controlling its expression. (ucl.ac.uk)
  • Current chemotherapy, such as gemcitabine or 5-fluorouracil, coupled with radiotherapy may improve the quality of life of some patients, but their survival benefit is very limited [ 4 ]. (oncotarget.com)
  • Luciferase catalyzes the oxidation of a substrate, leading to the emission of photons. (addgene.org)
  • The region of an enzyme that interacts with its substrate to cause the enzymatic reaction. (lookformedical.com)
  • Furthermore, a series of partial deletions of the 5′-flanking region illustrated that both the proximal and distal clusters of PXREs are required for maximal induction of rat oatp2 by PCN. (aspetjournals.org)
  • 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)
  • Furthermore, we analysed the 5'-flanking region of the human adipose differentiation-related protein ( adrp ) gene that responded to all subtypes of PPARs. (biomedcentral.com)
  • Recent genome-wide analysis has shown that DNA methylation spans long stretches of chromosome regions consisting of clusters of contiguous CpG islands or gene families. (biomedcentral.com)
  • Adenosine deaminase acting on RNA 1 (ADAR1) is closely related with circRNA formation through its adenosine-to-inosine (A-to-I) RNA editing, which decreased circRNAs production by lowering the complementarity of circRNA precursor's flanking introns [ 13 , 14 ]. (biomedcentral.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)
  • Firefly luciferase is considerably brighter than Renilla luciferase, but is more susceptible to enzyme inhibition. (addgene.org)
  • Innate immune and inflammatory responses are normal self-protection mechanism that eliminate pathogens and resist microbial invaders ( 5 , 6 ). (frontiersin.org)
  • Analysis of transcription factor (TF) and ChIP- seq datasets identified overlap of 70 SNPs with TF binding regions of relevant lung cell lines (e.g. (cdc.gov)
  • Metastatic burden was examined by quantitative PCR, through amplification of a region of the luciferase gene from lung DNA. (biomedcentral.com)
  • A southern blot using hybridization of gag, pro, env derived probes revealed a complex distribution of HERV-Ws in the human haploid genome with 70 gag, 100 pro, and 30 env regions. (wikipedia.org)
  • But, it is unclear about signal transduction pathway and gene regulatory region of human betadefensin-2 (hBD-2). (ajou.ac.kr)
  • This effect of haem was subsequently shown to be dependent on the -6.3 to -3.5 kb region of the 5'-flanking region of cALAS-1 and requires the native orientation of the region. (umassmed.edu)
  • We first identified the transcription start site of cNANOG by 5′-rapid amplification of cDNA ends PCR analysis. (biomedcentral.com)
  • 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)
  • 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)
  • This effect is mediated through specific transcription factor binding within the -52/+118 region of the PDGF alpha R gene. (nih.gov)
  • 5 , 6 In concordance with the role of nodal cilia in L-R asymmetry, mutations in the motor proteins Kif3A and Kif3B, and the transcription factor Foxj1 in the mouse have demonstrated that ciliary motility is important for L-R asymmetry. (bmj.com)
  • In addition, four other factors without consensus binding sites in this region were also recruited, possibly as components of transcription factor complexes. (elsevierpure.com)
  • In all, our studies identify thousands of breast tumor hypermethylated regions for the first time, in particular, discovering seven large contiguous hypermethylated gene clusters. (biomedcentral.com)