• About half of human genes use alternative cleavage and polyadenylation (ApA) to generate messenger RNA transcripts that differ in the length of their 3' untranslated regions (3' UTRs) while producing the same protein. (nih.gov)
  • Regulation of gene expression is primarily controlled by changes in the proteins that occupy genes' regulatory elements. (nature.com)
  • Genes are commonly generate isoforms from the same locus to produce different mRNA and thus give rise to different length of amino acid with similar protein functions. (hypothes.is)
  • The protein or DNA sequence of these genes, Lst1 and SEC24C, are highly similar and evolutionary share the common ancestor, termed as homolog. (hypothes.is)
  • Non-protein coding genes (e.g. rRNA genes, tRNA genes) are not translated into protein. (wikidoc.org)
  • TETRASPANIN (TET) genes encode conserved integral membrane proteins that are known in animals to function in cellular communication during gamete fusion, immunity reaction, and pathogen recognition. (unical.it)
  • NUCLEAR LOCALIZATION SIGNALGREEN FLUORESCENT PROTEIN/b-GLUCURONIDASE reporter lines throughout the life cycle, which predicted functional divergence in the paralogous genes per clade. (unical.it)
  • Fluorescence microscopy is extensively employed in zebrafish research due to the availability of fluorescent reporter genes and fluorophore-labeled probes. (bioimager.com)
  • Upright or boom stand fluorescence microscopy is widely used for in vivo imaging of zebrafish due to the availability of fluorescent reporter genes and fluorophore-labeled probes. (bioimager.com)
  • Overexpression of the RNA-binding protein HuR (ELAVL1) occurs during colon tumorigenesis and is abnormally present within the cytoplasm, where it post-transcriptionally regulates genes through its interaction with 3'UTR AU-rich elements (AREs). (oncotarget.com)
  • At the molecular level, pitavastatin induced expression of the cyclin dependent kinase (CDK) inhibitor p21 in a cholesterol independent manner, blocked repressive phosphorylation of the Retinoblastoma tumor suppressor protein at CDK targeted sites, and reduced expression of E2F target genes required for progression through the G1/S boundary. (oncotarget.com)
  • The long 3' UTR of CD47 enables efficient cell surface expression of CD47 protein, whereas the short 3' UTR primarily localizes CD47 protein to the endoplasmic reticulum. (nih.gov)
  • 3' UTR-dependent protein localization has the potential to be a widespread trafficking mechanism for membrane proteins because HuR binds to thousands of mRNAs, and we show that the long 3' UTRs of CD44, ITGA1 and TNFRSF13C, which are bound by HuR, increase surface protein expression compared to their corresponding short 3' UTRs. (nih.gov)
  • The purpose of this technique is to analyze the activity of a gene transcription promoter (in terms of expression of a so-called reporter gene under the regulatory control of that promoter) either in a quantitative manner, involving some measure of activity, or qualitatively (on versus off) through visualization of its activity in different cells, tissues, or organs. (wikipedia.org)
  • Since there is no detectable GUS activity in higher plants, mosses, algae, ferns, fungi and most bacteria, the assay is ideally suited for gene expression studies in these organisms, and considered the reporter gene of choice for in plant science. (wikipedia.org)
  • Light output resulting from the luciferase reaction is captured using a luminometer and can be correlated with the amount of firefly luciferase protein produced and used to determine the activity of the promoter driving firefly expression. (thermofisher.com)
  • Importantly, a precise temporal description of RPRM protein expression during the development of neuronal structures is still missing. (frontiersin.org)
  • Moreover, zebrafish is very well-suited for systematic analysis of gene expression patterns at transcript level by whole-mount in situ hybridization (WISH), and at protein level by immunohistochemistry/immunofluorescence (IHC/IF). (frontiersin.org)
  • Herein we examine the temporal and spatial expression patterns of RPRM gene-products (mRNA and protein) during neural development. (frontiersin.org)
  • Note the close correlation between mRNA and protein expression dynamics. (elifesciences.org)
  • The respective protein dynamics were visualised with antibodies against the respective protein or GFP fusion protein ( C,F,I,L,N,P ). Note that although Mhc and Act88F show significant mRNA and protein expression already at 30 hr APF, this expression dramatically increases after 30 hr APF to reach even higher protein levels at 72 hr APF. (elifesciences.org)
  • The secondary reporter drives a steady level of expression of a different marker. (promegaconnections.com)
  • Green Fluorescent Protein (GFP) is a widely-used reporter with which to analyze protein localization and expression levels. (plantae.org)
  • Furthermore, the protein level is not affected by SA, indicating again that it is not due to expression-level effects. (plantae.org)
  • Gene expression is the process by which the inheritable information in a gene , such as the DNA sequence , is made into a functional gene product , such as protein or RNA . (wikidoc.org)
  • Several steps in the gene expression process may be modulated, including the transcription step and the post-translational modification of a protein. (wikidoc.org)
  • Because it is very difficult to clone a GFP-fused protein into its native location in the genome, however, this method often cannot be used to measure endogenous regulatory mechanisms (GFP-fusions are therefore most often expressed on extra-genomic DNA such as an expression vector ). (wikidoc.org)
  • Fusing a target protein to a reporter can also change the protein's behavior, including its cellular localization and expression level. (wikidoc.org)
  • Any step of gene expression may be modulated, from the DNA-RNA transcription step to post-translational modification of a protein. (wikidoc.org)
  • This finding was also supported by dual-luciferase transient expression assay and transient β-glucuronidase (GUS) reporter assay. (biomedcentral.com)
  • Genetic studies of osmoregulation in yeast have identified mucin-like proteins as critical regulators of osmosensitive signaling and gene expression. (prolekarniky.cz)
  • Fluorescence stereo microscopes can visualize specific molecules or structures in zebrafish tissues and cells, such as gene expression patterns, protein localization, and cellular processes. (bioimager.com)
  • It allows visualization of specific molecules or structures within live zebrafish, such as gene expression patterns, protein localization, and cellular processes. (bioimager.com)
  • Mechanistically, MS-444 disrupted HuR cytoplasmic trafficking and released ARE-mRNAs for localization to P-bodies, but did not affect total HuR expression levels. (oncotarget.com)
  • In vivo treatment of MS-444 inhibited HuR cytoplasmic localization and decreased COX-2 expression in tumors. (oncotarget.com)
  • Purified CyHV-2 ORF55 protein was obtained by prokaryotic expression, and the interacting peptide was screened out using phage display. (biomedcentral.com)
  • Reverse transcription quantitative PCR results demonstrated high expression of an actin-binding Rho-activating protein in the latter stages of virus-infected cells, and molecular docking, cell transfection and coimmunoprecipitation experiments confirmed that it interacted with the ORF55 protein. (biomedcentral.com)
  • Despite a lack of cellular localization, nuclear localization of GUS has been well observed. (wikipedia.org)
  • At the sub-cellular scale, maps document the spatial organization of proteins, RNA, DNA, and metabolites with nanometer precision and temporal acuity on the order of seconds. (stanford.edu)
  • We developed genomic locus proteomics (GLoPro), in which we combine CRISPR-based genome targeting, proximity labeling, and quantitative proteomics to discover proteins associated with a specific genomic locus in native cellular contexts. (nature.com)
  • To degrade the misfolded proteins and maintain the ER homeostasis, the cellular signaling will turn on the unfolded protein response to help the cell deals with problematic proteins for recycle and degradation. (hypothes.is)
  • The most commonly used method is to perform a Western blot against the protein of interest, whereby cellular lysate is separated on a polyacrylamide gel and then probed with an antibody to the protein of interest. (wikidoc.org)
  • Cellular and subcellular immunohistochemical localization and quantification of cadmium ions in wheat (Triticum aestivum). (cdc.gov)
  • Several kinds of GUS reporter gene assay are available, depending on the substrate used. (wikipedia.org)
  • The GUS assay, as well as other reporter gene systems, can be used for other kinds of studies other than the classical promoter activity assay. (wikipedia.org)
  • By proximity ligation assay (PLA), we demonstrated that the mCherry-Arc fusion protein retains the Arc function by interacting with the transmembrane protein stargazin in postsynaptic spines. (lu.se)
  • Another post describes how to choose the best dual-reporter assay for your experiments. (promegaconnections.com)
  • The underlying mechanisms of action of LIPH-4 were explored through bioinformatics, luciferase reporter assay, RNA-immunoprecipitation assay and immunoblot. (researchsquare.com)
  • Protein and peptide interaction prediction and dot-blot overlay assay confirmed that peptides identified by phage display could interact with the ORF55 protein. (biomedcentral.com)
  • We found that one suppressor, the transmembrane protein PTR-23, is co-expressed with osm-8 in the hypodermis and strongly suppresses several Osm-8 phenotypes, including the transcriptional activation of many osmosensitive mRNAs, constitutive glycerol accumulation, and osmotic stress resistance. (prolekarniky.cz)
  • Our studies are the first to show that an extracellular mucin-like protein plays an important role in animal osmoregulation in a manner that requires the activity of a novel transmembrane protein. (prolekarniky.cz)
  • The herpes simplex virus type 1 (HSV-1) glycoprotein N (gN/UL49.5) is a type I transmembrane protein conserved throughout the herpesvirus family. (mdpi.com)
  • This process is analogous to hydrolysis of X-gal by Beta-galactosidase to produce blue cells as is commonly practiced in bacterial reporter gene assays. (wikipedia.org)
  • Cell-based and in vitro assays show that Retro-2 blocks delivery of newly-synthesized TA-proteins to the ER-targeting factor ASNA1 (TRC40). (elifesciences.org)
  • Previously, we described some of the advantages of using dual-reporter assays (such as the Dual-Luciferase® , Dual-Glo® Luciferase and the Nano-Glo® Dual-Luciferase® Systems). (promegaconnections.com)
  • While there are many advantages to dual-reporter assays, they require careful planning to avoid common pitfalls. (promegaconnections.com)
  • Here we show in human cell lines that alternative 3' UTRs differentially regulate the localization of membrane proteins. (nih.gov)
  • Increased ion accumulation raises cytoplasmic ionic strength, which can disrupt protein structure and function [3] . (prolekarniky.cz)
  • Moreover, they infect cells with viruses, or express individual viral proteins, to follow the virus's actions and the host cell response. (the-scientist.com)
  • In the zebrafish, rprma mRNA is expressed in the olfactory placodes (OP) and olfactory epithelium (OE), rprmb is observed in the tectum opticum (TeO) and trigeminal ganglion (Tg), whereas rprml is found primarily in the telencephalon (Tel). At protein level, RPRM is present in a subset of cells in the OP, and neurons in the OE, TeO, hindbrain and sensory peripheral structures. (frontiersin.org)
  • When inducing long-term potentiation (LTP) in the hippocampus, we observed an increase of Arc protein highly correlated with an increase in fluorescent intensity and the number of mCherry-positive cells. (lu.se)
  • These small highly conserved actin-binding proteins are essential regulators of actin dynamics in living cells. (rupress.org)
  • GFP fusion gene has been fully sequenced, its fluorescence confirmed and its function tested in HEK293 cells coexpressing an NF-κB reporter plasmid ( pNiFty-SEAP ). (invivogen.com)
  • Insect cells have been central to many scientific advances, being utilized in recombinant protein, baculovirus, and vaccine and viral pesticide production, among other applications (5). (promegaconnections.com)
  • But, how do you pick the best way to get the vector into your cells to express the reporter? (promegaconnections.com)
  • A series of interconnected protein and carbohydrate networks that surround and support cells in 3 dimensions. (hypothes.is)
  • To determine the mechanisms leading to actin remodeling, fluorescent protein fusions of the BSMV triple gene block (TGB) proteins were coexpressed in cells with the actin marker DsRed: Talin. (ppjonline.org)
  • To track differentiating cell populations, reporter cell lines generated by homologous recombination (knock-in) or via BAC transgenes have been widely used in mouse cells (5-8 ). (lu.se)
  • Intracellular transport of plant viruses during infection operates via unique interactions between viral proteins and selected host cytoskeletal and membrane elements. (ppjonline.org)
  • During viral infection, the ORF55 protein exerts its biological function through interactions with host proteins. (biomedcentral.com)
  • There are a variety of ready-to- transduce viral vector collections including constitutively expressed or inducible fluorescent proteins for nuclear or membrane localization and bioluminescence reporters for 3D drug screening. (sbpdiscovery.org)
  • Reporter systems have been used for the determination of the efficiency of gene delivery systems, the intracellular localization of a gene product, the detection of protein-protein or protein-DNA interactions, the efficiency of translation initiation signals and the success of molecular cloning efforts. (wikipedia.org)
  • These instrumentation-based innovations have occurred in parallel with the development of new molecular approaches to label protein targets, to evolve new biosensors and to target them to appropriate cell types or subcellular compartments. (bordeaux-neurocampus.fr)
  • Nonetheless, 2009 data has shown for that to start with time that the subcellular localization selleck chemicals of Kaiso while in the cytoplasm of the cell is straight connected using the bad prognosis of patients with lung cancer, and close to 85 to 95% of lung cancers are non small cell. (screeninglibraries.com)
  • Kaiso and prognosis As expected for a transcriptional aspect, the Kaiso Inhibitors,Modulators,Libraries protein is often discovered while in the nucleus of quite a few tumor or non tumor derived mammalian cell lines. (screeninglibraries.com)
  • Use a primary and secondary reporter vector transiently transfected into your favorite mammalian cell line. (promegaconnections.com)
  • The lilli gene encodes a nuclear protein related to the AF4/FMR2 family. (sdbonline.org)
  • Although highly sensitive, LacZ/X-gal staining is not quantitative, and does not provide the high resolution possible with fluorescent or bioluminescent proteins. (vectorbuilder.com)
  • To enable the tracking of Arc molecules from individual neurons in vivo, we devised an adeno-associated virus (AAV) mediated approach to tag the N-terminal of the mouse Arc protein with a fluorescent reporter using CRISPR/Cas9 homologous. (lu.se)
  • To enable the tracking of Arc molecules from individual neurons in vivo, we devised an adeno-associated virus (AAV) mediated approach to tag the N-terminal of the mouse Arc protein with a fluorescent reporter using CRISPR/Cas9 homologous independent targeted integration (HITI). (lu.se)
  • When To Use Fluorescent Reporter, Luciferase Or LacZ? (vectorbuilder.com)
  • Our results indicate that actin cytoskeleton interactions are important in BSMV cell-to-cell movement and for CW localization of TGB3. (ppjonline.org)
  • This fusion protein can be used to study the localization of TLR1. (invivogen.com)
  • The drug selectively inhibits the kinase activity of the BCR ABL fusion protein. (screeninglibraries.com)
  • When newly synthesized protein is not properly folded into their correct orientation in the endoplasmic reticulum (ER), the ER now is under the stress to degrade the misfolded protein. (hypothes.is)
  • Fluorescent proteins are much brighter than luciferase, which benefits many types of experiments, but in tissue samples or live animals, background, autofluorescence, and light scattering can make fluorescent proteins problematic. (vectorbuilder.com)
  • In our model of 3' UTR-dependent protein localization, the long 3' UTR of CD47 acts as a scaffold to recruit a protein complex containing the RNA-binding protein HuR (also known as ELAVL1) and SET to the site of translation. (nih.gov)
  • TLR1-GFP fusion gene was generated by fusing the green fluorescent protein (GFP) gene to the C terminus of human Toll-like receptor 1 (TLR1) gene. (invivogen.com)
  • Another commonly used method for assaying the amount of a particular protein in a cell is to fuse a copy of the protein to a reporter gene such as Green fluorescent protein , which can be directly imaged using a fluorescent microscope. (wikidoc.org)
  • For FPs, one often compares the MB to that of enhanced Green Fluorescent Protein (EGFP), which is about 30000 M -1 .cm -1 . (ibs.fr)
  • The primary reporter is commonly used as a marker for a gene, promoter, or response element of interest. (promegaconnections.com)
  • Atg8 protein is a marker protein to observe autophagosome formation. (hypothes.is)
  • Using the endosome marker Rab5, we observed localization into endosomes. (mdpi.com)
  • DiX-indigo, can associate with lipids to diffuse far from the site of enzyme activity, which shows a lack of cytosolic localization and irregularity of substrate penetration. (wikipedia.org)
  • This appears to be uncommon because Kaiso features a signal NLS really conserved and demanded for any protein with nu clear localization. (screeninglibraries.com)
  • Simultaneous immunoassay of phosphorylated proteins based on apoferritin templated metallic phosphates as voltammetrically distinguishable signal reporters. (cdc.gov)
  • A major modulator of actin filament disassembly is the actin-depolymerization factor/cofilin family of proteins (hereafter referred to as cofilin). (rupress.org)
  • Severing increases the number of actin filament pointed ends and, in conjunction with capping of barbed ends by capping proteins, stimulates filament disassembly ( Andrianantoandro and Pollard, 2006 ). (rupress.org)
  • This localization was found to be unaffected by manipulations that were expected to change polymerization rates at the leading edge, which should alter the relative abundance of ATP-, ADP-Pi-, and ADP-actin subunits within the filaments. (rupress.org)
  • We propose that during translation the scaffold function of 3' UTRs facilitates binding of proteins to nascent proteins to direct their transport or function--and this role of 3' UTRs can be regulated by ApA. (nih.gov)
  • Right: Illustrative neuronal targets consist of glutamate receptors, adhesion protein complex and scaffold proteins (dashed lines represent the non-resolved part of the complexes) represented in the confined environment of the synaptic cleft (~20-25 nm). (bordeaux-neurocampus.fr)
  • CD47 protein localization occurs post-translationally and independently of RNA localization. (nih.gov)
  • Misfolded protein are retained in the ER and subjected for proteosomal degradation which occurs in the cytosol. (hypothes.is)
  • The GUS system is not the only available gene reporter system for the analysis of promoter activity. (wikipedia.org)
  • The activity-regulated cytoskeleton-associated (Arc) protein is essential for synaptic plasticity and memory formation. (lu.se)
  • To image affinity probes, these are generally further functionalized with particles (not shown, from ~5 nm to a few tens of nanometres), fluorescent proteins or organic dyes. (bordeaux-neurocampus.fr)
  • Organic osmolytes accumulation is mediated by metabolic biosynthesis or transport into the cell via specialized organic osmolyte transporter proteins [5] . (prolekarniky.cz)
  • Affinity probes can either be developed against any protein target or be specific for a molecular target (specified in parentheses). (bordeaux-neurocampus.fr)
  • These observations led to the hypothesis that cofilin localization is largely dependent on accessory factors or other regulatory inputs, such as phosphorylation or pH. (rupress.org)
  • Stochastic Optical Reconstruction Microscopy, STORM , is one of a family of Nobel Prize winning super-resolution Single Molecule Localization Microscopies ( SMLM ) for the visualization of biological systems with an optical resolution measured in the tens of nanometers (nm) in the x, y, and z directions. (microscopyu.com)
  • Given that mucins and transmembrane proteins play similar roles in yeast osmoregulation, our findings suggest a possible evolutionarily conserved role for the mucin-plasma membrane interface in eukaryotic osmoregulation. (prolekarniky.cz)
  • Here, we show that NEMO phosphorylation by GSK-3β leads to NEMO localization into multivesicular bodies (MVBs). (mdpi.com)
  • Recent studies utilizing immunohistochemistry evaluation of ordinary and tumor tissue unveiled that Kaiso protein is predominantly localized inside the cytoplasm in the cell or is totally absent, though. (screeninglibraries.com)
  • 1 probable explanation is the fact that Kaiso, like other proteins or variables that normally reside during the cytoplasm, need a post translational modification, to get targeted and translocated towards the cell nucleus. (screeninglibraries.com)
  • glycerol, sorbitol, myo-inositol, etc), which are non-ionic solutes that can be accumulated to high (100s of mM) concentrations without affecting cell structure and/or protein function [4] . (prolekarniky.cz)
  • The serine/threonine kinase protein kinase D1 (PKD1) is a stress-responsive kinase and sensor for reactive oxygen species, which can initiate cell survival through NF-κB signaling. (biologists.com)
  • 2440 sequence verified ORFs from the Ultimate ORF Collection (LT) are available as sub-libraries including secreted protein (858 ORFs), apoptosis (476 ORFs), kinase (373 ORFs), and innate immunity (172 ORFs). (sbpdiscovery.org)
  • This hybrid protein absorbs blue light (major peak at 480 nm) and emits green light (major peak at 505 nm). (invivogen.com)
  • As shown on the figure (so called Jablonski diagram), when a molecule (e.g. a fluorescent protein) in the electronic ground state absorbs a photon of suitable wavelength, it accesses the first electronically excited state. (ibs.fr)
  • The barrel structure produces a very tight packing of the interior of the protein, so that the chromophore and water molecules are largely fixed into place by hydrogen bonds, with limited (but not impossible) diffusion of ions or small molecules. (ibs.fr)
  • The Arc gene, which contains remnants of a structural GAG retrotransposon sequence, produces a protein that self-assembles into capsid-like structures harboring Arc mRNA. (lu.se)
  • While nine spCas9 gene editing sites surround the Arc start codon, the accuracy of the editing was highly sequence-dependent, with only a single target resulting in an in-frame reporter integration. (lu.se)
  • A foreign protein is first expressed in fusion with one of the phage capsid proteins by cloning the foreign DNA sequence in frame with the gene encoding the chosen capsid protein [ 22 ]. (biomedcentral.com)
  • In many cases this can limit reproducibility or quantitation of experiments, when compared to fluorescent proteins. (vectorbuilder.com)
  • Fluorescent proteins function by absorbing light of one color (excitation), and then emitting lower-energy light of a different color (emission). (vectorbuilder.com)
  • Finally, we recorded mCherry-Arc interaction with presynaptic protein Bassoon in mCherry-negative surrounding neurons at close proximity to mCherry-positive spines of edited neurons. (lu.se)
  • CRISPRi genetic interaction analysis revealed Retro-2 activity resembles disruption of the transmembrane domain recognition complex (TRC) pathway, which mediates post-translational ER-targeting and insertion of tail-anchored (TA) proteins, including SNAREs required for retrograde transport. (elifesciences.org)
  • The GUS reporter system (GUS: β-glucuronidase) is a reporter gene system, particularly useful in plant molecular biology and microbiology. (wikipedia.org)
  • One of the most interesting features of the GFP chromophore is that it does not require a catalytic system to be formed, but is autocatalytically assembled after protein production, requiring only molecular oxygen as a cofactor for its full maturation. (ibs.fr)
  • After maturation, these three amino-acids form a para-HydroxyBenzylideneDimethylImidazolinone (p-HBDI) structure, responsible for the light absorption and emission properties of the protein. (ibs.fr)
  • Mechanical forces are transduced into biochemical signals by specialized proteins. (nature.com)
  • Fluorophores for STORM and SMLM include synthetic dyes, fluorescent proteins (FPs), and even Quantum Dots (QDots). (microscopyu.com)