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  1. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ... plasmids expressing luciferase Luciferase reporter assay The typical luciferase-containing plasmid is...firefly luciferase was published in 1985, several studies utilized luciferase as a genetic reporter in plant...and a luciferase gene (one of many available to choose from) is now a common feature in reporter plasmids...Learn How Luciferase Can Be Used In Concert with Fluorecent Proteins in Nano Lanterns Reporter gene assays...assays A reporter gene, such as luciferase, usually serves as an indicator of transcription within cells...Well-established Biological sources of luciferase Although luciferase isolated from the firefly beetle (Photinus... most frequently used bioluminescent reporter, other luciferases have been identified in various species...
  2. Technologies Enabled by NanoLuc® Luciferase

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    Blog Post
    ...researchers took a FRET reporter and adapted it into a BRET reporter by using NanoLuc® Luciferase to excite the ...codon-optimized Firefly luciferase genes (e.g., luc2), and NanoLuc® Luciferase are available from the ... creation of NanoLuc® Luciferase from a rather dull 19kDa Oplophorus luciferase by Promega’s Advanced ...than our more common luciferases like firefly (FLuc) and Renilla (RLuc) luciferase. This is important not...has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged...furimazine substrate for NanoLuc luciferase. Both GpNLuc and OgNLuc BRET reporters produced bright signals without...excited by NanoLuc® luciferase (NLuc) and set about to create an in vivo imaging reporter by linking two CyOFP1...
  3. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

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    Blog Post
    ...Administration of Substrate: Some luciferase reporters, such as Renilla luciferase (RLuc) or NanoLuc, require... imaging tools The luciferase reporter gene The green fluorescent protein reporter gene Resources on ...developed in vivo bacterial reporters that use fluorophores and luciferases to track bacteria in real time... time, but each of these reporters has its drawbacks. Acoustic reporter genes (ARGs) overcome these limitations...existing bacterial reporters Limited Depth of Signal/Imaging: Fluorescent reporters are not as well suited...fluorescent or bioluminescent reporters usually only detects the signal of the reporter and fail to pinpoint what...signal is coming from.   Acoustic reporter genes (ARGs) Acoustic Reporter Genes (ARGs) are engineered gene...
  4. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...to the widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have...fluorescent pH sensors, a set of luciferase-fluorophore pH fusion reporters, pRSETb-pHlash and pcDNA3.1+-... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...their reporter mRNAs. Using the TRICK plasmids, Halstead et al demonstrated that 91% of reporter mRNAs...expressing a separate reporter gene (EGFP, iRFP, IFP1.4, puromycin, neomycin or luciferase) to create a final...customized fluorescent reporters can be challenging enough, but incorporating multiple reporters into a single .... Nat Methods. 2011. High-throughput luciferase reporter system for detection of pre-mRNA splicing...
  5. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ... of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether it interacts...the promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...genes as reporters and some that repress instead of activate transcription. The HIS3 reporter system (... that avoid transcriptional reporters altogether; In the split-luciferase system, intein-mediated splicing...reconstitutes functional luciferase only when fusions of the two halves of the luciferase protein interact and...co-expressed in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing...referred to as the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI...
  6. Hot Plasmids: Fall 2024

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    Blog Post
    ...gene disruption in M. abscessus  Bioluminescent reporter of mechanical forces   Epitope tag antibody ...Western Blot for Anti-V5 [IPI-SV5-Pk1]. Addgene Report, https://doi.org/10.57733/addgene.l0ulu4. Hanke...doi.org/10.1128/jb.00335-23.  Bioluminescent reporting of mechanical forces with PILATeS By Emily P. ...mechanosensor called PILATeS, Pulling-Induced Luciferase Association Tension Sensor (Zhong et al., 2024...sensor STReTCh, the 11-amino acid BiT fragment of luciferase is embedded in the I10 domain of the human titin...tension. When this happens, BiT and the LgBiT luciferase fragment (expressed separately) assemble and ... reaction (Figure 5). The researchers used the reporter to visualize biologically relevant forces at focal...
  7. Luminescent Imaging with Nano-lanterns

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    Blog Post
    ...Renilla luciferase variant fused to an FP; when supplied with a luciferase substrate, the luciferase transfers...problems of low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently...Nano-lantern, a fusion of a mutagenized Renilla luciferase (RLuc8) with Venus. This luminescent technology...transfer (BRET). Photons emitted by a Renilla luciferase variant are used to excite the fused fluorescent...need for an excitation light source. Instead, a luciferase substrate like coelenterazine is supplied in ...eNL constructs use the brightest characterized luciferase, NanoLuc, and its substrate furimazine. In addition...binding. Suzuki et al. also envision eNLs as good reporter genes for endogenous protein fusions created using...
  8. Luciferase Plasmid Collection

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    Collection
    ...types of luciferase found in Addgene’s collection are Firefly luciferase (Fluc), Renilla luciferase (Rluc...Renilla luciferase in plants Diego Orzaez 118069 MLRV Multiple Multiple Multi-luciferase reporter vector...collection of luciferase plasmids for assays of transcriptional regulation and bioluminescent reporters. Plasmid...Plasmid Collections Luciferase Plasmids Luciferase Plasmid Collection You may also like... Fluorescent Proteins...Proteins Blog: Luciferase 101 Blog: Technologies Enabled by NanoLuc® Luciferase Blog: Luminescent Imaging...Empty Backbones Expression Constructs Reporter Constructs Luciferase is the enzyme responsible for the bioluminescence...photobleaching has made luciferase a common choice in assays ranging from use as a reporter gene in vitro and...
  9. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ...replaced by a single (non-viral) reporter gene (e.g. firefly luciferase or eGFP) flanked by the EBOV-specific...as a template to transcribe the reporter gene (e.g. firefly luciferase). The polymerase recognizes the...viral infection), producing firefly luciferase. Firefly luciferase can readily be measured to quantify...necessary to direct minigenome replication and reporter gene transcription by the EBOV polymerase (reviewed...start signal and initiates transcription of the reporter gene. The polymerase will elongate the mRNA until...
  10. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ... Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance... editing. eGFP reporters for single base editing by APOBEC-Cas9 Though the ACE reporter quantifies APOBEC-Cas9...DSB in their previous reporter, the Harris Lab created a panel of eGFP reporters to quantify the on-target...editing reporters. Image from St. Martin et al., 2019. The Harris lab used these eGFP reporters to analyze...Transient reporter for editing enrichment (TREE) The Wang and Brafman lab developed a transient reporter for...cells segregated solely by a reporter of transfection, which only report the efficiency of plasmid delivery... functional reporter system to identify and enrich for base-editing activity The reporters mentioned above...
  11. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ... transformation. Choose good reporter genes There are many great reporter genes available and which one...confirm transformation success. Luciferases are great quantitative reporters and use a substrate to produce...the T-DNA region with our gene of interest or a reporter gene so that the transformation and gene expression...Once we were on the right track, we switched to luciferases to more easily quantify the effect of the different...different treatments on transformation success. Some reporter gene examples are below. The GUS gene (β-glucuronidase...histochemical GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate...produce bioluminescence. Luciferases usually have shorter mRNA and protein half-lives then GUS and are therefore...
  12. SARS-CoV-2 Pseudotyped Virus

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    Collection
    ... Lentiviral reporter vector expressing firefly luciferase. pCMV-FLuc - Retroviral reporter vector expressing...expressing firefly luciferase and ZsGreen. NL4-3 mCherry Luciferase - Lentiviral dual reporter vector expressing...EGFP. Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing firefly luciferase with neomycin...Envelope and Packaging Plasmids Reporter Plasmids A few examples of reporter plasmids that can be used for...expressing firefly luciferase. pHAGE-CMV-Luc2-IRES-ZsGreen-W - Lentiviral dual reporter vector expressing...expressing mCherry and firefly luciferase. pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-Hygro - Lentiviral...
  13. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ...cultured cells and activate expression of a luciferase reporter. CPP was not required for protein entry into...cells! After verifying the specificity of their luciferase assay, they concluded that components in the ...primary cell types using Cre and various loxP reporters. In both mouse and human embryonic stem cells,...Plasmids @Addgene: iTOP-tested proteins and reporters Previous Protein Delivery Techniques: Introduction...
  14. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ... and a UAS-driven luciferase reporter construct. In the absence of GFP, no reporter output was observed... and a reporter construct into the mouse retina. GFP expression robustly activated the reporter gene TdTomato...and Cre-DOG, respectively, repurpose popular GFP reporter lines for more sophisticated experimental manipulations...create a complete transcription factor, activating luciferase transcription. Tang et al. coined this system...GFP. They then tested T-DDOGs with two mouse GFP reporter lines; again, TdTomato was seen only in cells ...tested in retinal electroporation studies, DsRed reporter expression was induced in ~76% of GFP+ cells, ...
  15. Bacterial Expression Systems

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    Collection
    ...Tagging Purification Controlled Expression Reporter Plasmids Reporter Plasmids Tagging and Visualization Purification...expression at a specific level. Return to Top Reporter Plasmids Reporter plasmids can be used to detect events...they activate expression of the reporter gene. Plasmid ID Promoter Reporter PI Purpose pRU Series Various...pTac GFP, Luciferase (LuxCDABE) Attila Karsi Broad host range vectors for expressing luciferase and fluorescent...Purification Controlled Expression Reporter Plasmids You may also like… Plasmids 101 Addgene's Molecular...promoters, their associated transcription factors, and reporter genes. pCS-PesaRlux 47640 PesaR 3OC6HSL (Quorum...Quorum sensing molecule) Cynthia Collins Contains luciferase inducible by the quorum sensing molecule, 3OC6HSL...
  16. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    ...light include chemiluminescent proteins, like luciferases, as well as fluorescent ones, like Green Fluorescent...responsible for encoding GFP, and, in 1992, he reported its sequence. Soon after that, in 1994, Prasher...he lived! His hunch paid off, and he was able to report new fluorophores like DsRed from Discosoma sp. ...
  17. SunTag and Fluorescent Imaging

    Type
    Blog Post
    ...protein function, thereby creating a single-molecule reporter of intracellular processes. Initially, Tanenbaum...Visit Our Fluorescent Protein Guide Page Find Luciferase Plasmids ...
  18. Plasmids 101: Multicistronic Vectors

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    Blog Post
    ...along with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to ... using IRES elements. Further, scientists have reported lower expression of the downstream cistron due...eukaryotic cells, and, although some scientists report close to 100% cleavage with some of these peptides...that already has the multicistronic element and reporter cloned in. In these plasmids you would simply ... 2A element (depending on the placement of the reporter gene). Addgene's collection offers a variety of...Retroviral pEF1a-IRES-Neo IRES Mammalian MSCV-IRES-Luciferase IRES Retroviral pWPI IRES Lentiviral AmCyan-P2A-mCherry...multicistronic element(s), or you could replace one of the reporter genes above with your gene or genes of interest...
  19. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ...expression plasmids, gene knock-down plasmids, reporter plasmids, viral plasmids, and genome engineering...Green Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor the level of a chemical in a particular...
Showing: 1 - 20 of 39 results