...Fluorecent Proteins in Nano Lanterns Reporter gene assays A reporter gene, such as luciferase, usually serves... is also the most commonly used bioluminescent reporter. This beetle emits a yellow-green light with a...several studies utilized luciferase as a genetic reporter in plant and mammalian cells. Luciferase assays...available to choose from) is now a common feature in reporter plasmids.  Learn How Luciferase Can Be Used In...transcription within cells, where detection of the reporter protein or its enzymatic activity is measured....expression can be determined by detection of the reporter in a specific assay, which ideally would have .... Production of photons by this bioluminescent reporter occurs slower than fluorescent-based methods, ...

... registered reports, the gold standard for testing hypotheses Finally, Registered Reports combine all ...Reproducibility Project: Cancer Biology, where reported methods may not include how many tests were conducted...attention on the results instead of the methods. Unreported flexibility and questionable research practices...analysis code, and other materials. The use of reporting guidelines can also help remind us to include ...important times to measure that growth rate, you report the results of all four of those tests and not ...fifth time has unpredictably high rates, should be reported as a new discovery that deserves confirmation ...best practices of Open Science. With a Registered Report, peer review occurs before results are known. The...

...invert the reporter and GeneR fragment. Then the second recombination event would excise the reporter from ...Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters are delivered using AAV injection...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ...Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...shown in Figure 2, the cassette should contain a reporter DNA sequence (such as GFP) in the sense orientation...the two pairs of target sites. By flanking the reporter with one loxP site and one lox511 site in the ...would ensure that the reporter remains silent until a cell or tissue-specific Cre is provided. Since the FLEx...

...fundamentally the reporter construct monitors the expression of a nearby gene. The reporter gene serves as... screens to introduce reporters throughout an organism’s genome. These reporters have historically been...stimulate the expression of the reporter. The system is made up of a reporter gene flanked by a minimal promoter...promoter – all rely on the random integration of a reporter construct in close proximity to an endogenous ...example, if green fluorescent protein (GFP) was the reporter gene and green cells were observed in the kidneys... for with an uncharacterized gene. The actual reporter construct can be something phenotypic (such as...promoter. The system is composed of a promoter-less reporter which needs to integrate within an exon of a gene...

... transformation. Choose good reporter genes There are many great reporter genes available and which one...the T-DNA region with our gene of interest or a reporter gene so that the transformation and gene expression...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...transformation success. Luciferases are great quantitative reporters and use a substrate to produce bioluminescence...half-lives then GUS and are therefore good temporal reporters to measure promoter activity, for example. Useful...Agrobacterium. This is especially problematic with reporter genes as you might observe expression from Agrobacterium...

... versions of this reporter to target the K9 (pcDNA3-K9 histone methylation reporter) and K27 (pcDNA3-K27...The K9 reporter uses the HP1 chromodomain to recognise lysine 9 methylation, while the K27 reporter uses... methylation in living cells. Essentially this reporter construct consists of one substrate (the histone-derived...-K27 histone methylation reporter) positions on histone H3, which are known to be involved in repression...methylation of these sites was controlled, the reporter consistently showed significantly different FRET...states. Thus, this system is a reliable tool in reporting methylation status of these sites. This new technology...Alice Y. Ting. "Genetically encoded fluorescent reporters of histone methylation in living cells." Journal...

...technology to modify three different sites in the EGFP reporter gene (Fu et al., Nat Biotechnol. 2013). The expression...

...ACE) in real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry...and N177F mutations). Its excitation maximum is reported as 603 nm, while the emission maximum is 675 nm... ACE system: A versatile in vivo fluorescent reporter for rapid quantification of base-editing and Cas9...editing tools, the Reuben Harris lab has deposited a reporter plasmid that allows for rapid fluorescence-based... out a frame-shift mutation within the mCherry reporter. Quantification of editing efficiencies simply...containing rat APOBEC1. Finally, they show that the ACE reporter can also quantify Cas9 cleavage, thus exhibiting...editing activities. Click here to find the ACE reporter plasmid! Martin et al., Nucleic Acids Research...

...systems. Jump to Table: Cloning and Genomic Tools Reporters and Tags Library Construction Integration Recombination...

...random variation, or noise, in reporter expression. To do so, one places reporter expression under the control...expression of a reporter protein, it is possible to change the amount of noise in reporter gene expression...deliver the target antibody. TRIM-Away reagents and reporters are available from Addgene - we hope they will...lab has deposited plasmids encoding fluorescent reporters and other proteins controlled by GAL4 and ZIF268...progesterone controlled transcription factor as well as reporters specific to each transcription factor. They then... of estradiol and progesterone and showed that reporter expression is dependent only upon its cognate ...

...fluorescent reporter, directly couple silencing of your gene target to silencing of the reporter. This is...fusing your gene target sequence to a fluorescent reporter sequence (discussed below). Examples of these ...contains a target sequence against the fluorescent reporter in an inverted repeat orientation. Therefore, ...sequence against your gene(s) of interest and the reporter. This approach greatly enhances experimental throughput... observable phenotypes quantified. Fluorescent reporter-based RNAi constructs have been designed and widely...processes to isolate RNAi plants. With a visual reporter of RNAi, phenotypes, such as morphological parameters...identified as undergoing RNAi through the fluorescent reporter and manually extracted. This process is tedious...

...unique skills and strengths (you will likely have reports with different levels of experience), find out ...tasks before I was a manager! Second, my direct report was not engaged because I only left him/her with...you’re delegating a recurring task to a seasoned report, there should be questions! Remember that delegating...itself is counter-productive. I always assume that reports want to succeed at work and providing feedback ...feedback is needed for that. In many cases reports are not aware of issues until you bring them up. I wouldn’...wouldn’t go as far as saying that your reports will be grateful for the feedback, but at least you can start...someone’s hard work and reward a job well done. Your report will (needs to) feel valued. Talk to your manager...

...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...multicistronic element(s), or you could replace one of the reporter genes above with your gene or genes of interest...equivalent levels upstream of an IRES fluorescent reporter for easy detection.5  Just released: Addgene's...

...whole range of expression levels for these two reporters simultaneously. By fusing sfGFP and/or mCherry...

... people usually report? Addgene and other viral vector manufacturing facilities report the physical titer...cell transduction assays. Wild-type AAV2 has been reported to have a near-perfect physical-to-infectious ... However, for recombinant AAV2, the same study reported a physical-to-infectious particle ratio of 50:...(“full”) (Zeltner et al., 2010), while another reported that only 20% of recombinant AAV2 particles were...vector preparation protocol, and we are pleased to report that over 95% of viral capsids in representative...AAV aliquots. Our titer values (circled in red) report physical titer, which is measured by qPCR. ...nonsense to publication quality data." Scientific Reports 7 (2017). PubMed PMID: 28546538. PubMed Central...

...zebrafish Guide RNA expression plasmids for the EGFP Reporter Gene Guide RNA expression plasmids for endogenous...

...mutagenesis libraries or to insert heterologous genes, reporters, or other markers into a target genome. They did...Rep 2014 Jun 27;4:5469. MitoTimer plasmids to report mitochondrial turnover Mitochondrial health...Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-on system (pTRE-Tight-MitoTimer) to... to show its usefulness in cell culture to report changes in mitochondrial turnover and transport. pMitoTimer... Yan’s lab, which is constitutively expressed, reports on the balance of biogenesis and mitochondrial ...channelrhodopsins. CnChR1, nicknamed Chrimson, is the first reported yellow-peaked channelrhodopsin with a spectral... original APEX (enhanced ascorbate peroxidase) reporter from Alice Ting’s lab has enabled progress on ...

...molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can tell us about cellular events...higher apparent cellular brightness than previously reported widely used dimeric iRFPs (Table 1). In general...protein-protein interactions, we recommend using NIR split reporters based on miRFPs (Shcherbakova et al., 2016), since...fluorescence of co-transfected EGFP. e Originally reported as a monomer16, IFP2.0 was later found to be a...E., Sivanesan, D. & Michnick, S.W. An infrared reporter to detect spatiotemporal dynamics of protein-protein...Filonov, G.S. & Verkhusha, V.V. A near-infrared BiFC reporter for in vivo imaging of protein-protein interactions...evolution approach applied to fluorescent voltage reporters. Nat Chem Biol 14, 352-360 (2018). Yu, D. et al...

...a UAS-driven luciferase reporter construct. In the absence of GFP, no reporter output was observed in ... 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...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, ...

...for Endogenous Human Genes TALENs for the EGFP Reporter Gene Additional Plasmids compatible with the REAL...