...is typically used for gRNA. May contain reporter gene (e.g. GFP) to identify and enrich positive cells ...separate transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells....cloning in genome target sequence into EGFP reporter. Target DNA reporter system: Can evaluate component delivery...permanently knock out or knock-in a gene? Do you want to activate or repress gene expression? Are you trying to...Do you want to create a fusion with a reporter protein such as GFP? All of these outcomes can be most effectively...plasmids at Addgene that have been published as CRISPR testing tools: Traffic Light Reporter System: Can...conjunction with activator or repressor domains to control gene expression. Base editors can help you make precise...

... a Cre-dependent fluorescent CatCH-GFP reporter. Expression of GFP will only result in cells where Cre-mCherry...of the same transgene: an optogenetic reporter, channelrhodopsin, fused to a fluorescent GFP marker (CatCH-GFP... popular tool for gene delivery, but it has a drawback: how do you ensure your gene goes where you want...cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using phylogenetically conserved sequences...CatCH-GFP). The lab deposited AAV plasmids containing the active promoters at Addgene. Performance of ...projections. Generating animal models: Transgenic animals allow for the expression of genes in particular...Additional resources on the Addgene blog Read more about using AAV for retinal gene therapy Find more about...

...Chang-Deng Hu Green sfGFP Split super-folder GFP, our most-requested GFP(1-10) and GFP(11) Versatile protein...Commun. 2016 Bo Huang spGFP Split superpositive GFP Split-superpositive GFP reassembly is a fast, efficient...base FP (for example, EBFP2(1-10) can be used with GFP(11)). Some of the articles linked below only provide...Blue EBFP2 EBFP2(1-10) and Capri(1-10) for use with GFP(11) Multiplexed labeling of cellular proteins with... Kamiyama Cerulean Cerulean(1-10) for use with GFP(11) Multiplexed labeling of cellular proteins with... (abundant in eukaryotes) A Near-Infrared BiFC Reporter for In Vivo Imaging of Protein-Protein Interactions...green-yellow or orange-red fluorogen A split fluorescent reporter with rapid and reversible complementation. Tebo...

...-molecule reporter of intracellular processes. Initially, Tanenbaum et al. observed some GFP aggregation...scaffold fully occupied with GFP has a molecular weight of 1,400 kDa vs 24 kDa for GFP alone. Although traditional...aggregation, which they reduced by using superfolder GFP (sfGFP) with the small solubility tag GB1. In SunTag...nomenclature throughout the rest of this blog post, GFP refers to sfGFP-GB1.   Tanenbaum et al. examined the power... copies of the short epitope GCN4. GCN4 recruits GFP fused to the cognate scFV antibody, which is expressed...physiology. Having seen that mitochondrial tracker GFP-mitoNEET can impair mitochondrial function, they ...they examined the effects of mitoNEET-SunTag-GFP. As expected, they obtained bright images of mitochondria...

...endogenous gene. Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters are ...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ... the gene you would like to rescue with in the antisense orientation (let’s call the gene “GeneR”). The...rescue of a gene knockout: To test if the phenotype you are observing is really due to a single gene deleted... 2, the cassette should contain a reporter DNA sequence (such as GFP) in the sense orientation, followed...Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly... were designed as a genetic tool for researchers to conditionally manipulate gene expression in vivo using...

... is time consuming. GFP-tagged Atg8p or LC3: ATG8 is part of a group of genes that affect autophagy ...Resources on the Addgene Blog Beware of Fluorescent Protein Oligomerization Seeing Red: Simple GFP Photoconversion...Alkaline phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline phosphatase. Normally...with autophagosomal membranes, so tagging it with GFP allows for tracking the localization or accumulation...autophagosomes and autophagic bodies in yeast. A GFP-tagged version of LC3, the mammalian Atg8p homolog...pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic pHluorin (SEP). They are ...2008). Rosella: A fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in...

...elegans CRISPR/Cas9 for gene disruption in M. abscessus  Bioluminescent reporter of mechanical forces ...Anti-V5 [IPI-SV5-Pk1]. Addgene Report, https://doi.org/10.57733/addgene.l0ulu4. Hanke, T., Szawlowski,...Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126.   Gene disruption in...with expression levels comparable to GFP during early embryogenesis (Figure 3). mScarlet-I3 also offered...through the middle of each embryo are shown, comparing GFP, wrmScarlet (mScarlet), and the new mScarlet3 and...developed a novel dual-plasmid CRISPR/Cas9 system for gene disruption in Mycobacterium abscessus, an increasingly...Here, the researchers were able to target multiple genes in M. abscessus either simultaneously or stepwise...

...this general design scheme to generate two more OptoNBs, one against mCherry and one against GFP. Insertion...Lab monitored how AsLOV2 insertion into mCherry or GFP nanobodies affected binding of their target proteins...co-localization with a membrane-localized mCherry or GFP protein. With this system, if the OptoNB can bind....  AsLOV2 Inertion Site mCherry #1 mCherry #2 GFP Loop 1 dark-induced dark-induced dark-induced ...telephone, until the message reaches the nucleus where gene expression changes lead to cell proliferation and...activated Ras/Erk signaling by using a fluorescent reporter for Erk kinase translocation. These results demonstrate... protein activity Read about RANbodies, reporter nanobody fusions Learn more about using the secondary...

...pAAV.hSyn-FLEX.iGABASnFR.F102G pAAV-Syn-Archon1-KGC-GFP-ER2 pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2] pAAV-FLEX-tdTomato pAAV-GFAP104...pAAV-mDlx-NLS-mRuby2 CAG-NLS-GFP pAAV-hSyn-mCherry Read on to learn more!  Calcium sensor AAV The GFP-based calcium...pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2]) of this membrane-embedding...mCherry, pAAV-mDlx-NLS-mRuby2, CAG-NLS-GFP,and pAAV-hSyn-mCherry at Addgene. Viral vectors coming soon! These...37825  AAV8*, AAV9*  pAAV-CAG-GFP 50465  AAV8*, AAV9*  pAAV-hSyn-EGFP *Also coming in our new 20 ...with implanted electrodes. The Archon1 voltage reporter allows a way to monitor neuronal electrical activity...variant of jGCaMP7 in AAV1 at Addgene! See all calcium sensor AAV at Addgene. GABA sensor AAV GABA is the...

...released Anti-GFP [N86/8R] as another option for targeting GFP. But why might you need a second GFP antibody...on the Anti-GFP [N86/8R] page for more details. Image credit: Addgene.   Find anti-GFP [N86/8R] here...were compared head-to-head for gene expression changes of non-target genes and RNA-seq of a nuclear (MALAT1...PMCID: PMC9015817.   Meet anti-GFP [N86/8R]! by: Ashley Waldron Anti-GFP [N86/38.1R] was among the first...antibody?  Similarities:  Generated against full length GFP and converted to recombinant form by the NeuroMab...system Heat-inducible lentiviral Car constructs Anti-GFP [N86/8R] ready-to-use antibody Hyperfolder YFP ...ubiquitous promoter drives expression of an mCherry reporter.  After using mCherry for FACS sorting of the ...

...by replacing the genes on the T-DNA region with our gene of interest or a reporter gene so that the transformation...transformation. Choose good reporter genes There are many great reporter genes available and which one you...overexpressing a gene of interest in the plant. A common method for silencing genes Virus induced gene silencing...transformation success. Some reporter gene examples are below. The GUS gene (β-glucuronidase) with histochemical...Evaluation and comparison of the GUS, LUC and GFP reporter system for gene expression studies in plants." Plant...Agrobacterium. This is especially problematic with reporter genes as you might observe expression from Agrobacterium...Hikmet Budak. "Virus-induced gene silencing, a post transcriptional gene silencing method." International...

...live cells are GFP+. In this plot, with adjusted voltage, cells that did not receive the GFP protein are ...populations or a huge smear across your FSH vs reporter plot. In this post, I’ll walk you through how ...-case result. For example, you have introduced a GFP protein into your cells (visualized via the FITC ... to be clustered at signals below 103, while the GFP+ population cluster would be shifted to the right...samples and controls were processed the same way, the GFP- population should overlap with the negative control...control population (Figure 1). Based on where the GFP- population ends, you can draw a gate which selects...selects anything to the right of it, marking it as GFP+. Quantification for such graphs could simply be the...

...fuse them with a fluorescent reporter, such as GFP or mCherry, through genetic modification. As your protein...combination with that reporter, you can infer its presence through detecting the reporter's fluorescence. Another...previously, you can also use fluorescent reporters, such as GFP, and fluorophore-conjugated antibodies ...vector also encodes for eGFP, which serves as a transduction reporter. Note that eGFP is not fused to CD45...like APC-Cy7. Next comes our transduction reporter, eGFP. As eGFP emits fluorescence by itself and does not...overlap. Figure 4 compares the spectra of enhanced GFP (eGFP) and FITC, the latter being a commonly used fluorophore...combination of antibody-fluorophores, fluorescent reporters, and/or dye stains used in your flow cytometry...

...vitro and in vivo for gene expression and/or function. These data are sometimes reported or posted on the ... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, or both... Karl Deisseroth (Addgene viral prep # 55632-AAVrg). pAAV-CAG- FLEX-rc [Jaws-KGC-GFP-ER2] was a gift from... plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral helper sequences...recipient’s initial thaw will be accounted for in our reported titers. Lentiviral vectors are titered using a...copies of the lentiviral Rev responsive element (RRE) gene and the cellular ribonuclease P/MRP 30 kDa subunit...

...leader sequence. Using a variety of GFP and RFP fusions as their test genes, the researchers at the BIOFAB ...of varying strengths centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013...translational repressor to down-regulate a gene of interest. You can generate PUF proteins with a wide variety ...restriction enzymes and standardized primers to clone your gene of interest into both sequencing (pInitial) and ...abundance (meaning they are unlikely to be found in your gene of interest) and prevent incorporation of extensive...kit contain a variety of promoters to facilitate gene expression in bacteria, yeast, mammalian cells, ...various combinations of protein tags (His, Avi, and GFP) as well as protease sites to facilitate protein ...

...What are Acoustic Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used ...for visualizing gene expression in vivo.  But what ultrasound lacked was reporter genes that could produce...Shapiro Lab - developed Acoustic Reporter Genes (ARGs), which are sets of genes that, when expressed, enable...used to study gene expression in cells. But these reporters have limited use in vivo because they are based...vesicles gene clusters from a diverse set of gas vesicle-expressing species. The gas vesicle genes from Serratia...structural gene gvpA marked with EBFP2 and the second cassette contains the P2A-linked accessory genes gvpNJKFGWV...5209 (2017). Bourdeau, R. W. et al. Acoustic reporter genes for noninvasive imaging of microorganisms in...

...the addition of the enhanced GFP (EGFP)-type termini. mNeonGreen was reported as the brightest monomeric... to 3 times brighter than the most commonly used GFPs and YFPs. Its excitation maxima is at 506 nm and...Oregon created mTFP1 by transforming E. coli with a gene library of approximately 484 variants, including... or for live cell imaging of fusions and FRET reporters. mTFP1 is also insensitive to physiologically ... your research Subscribe to the Addgene blog Resources on Addgene.org Learn about the Michael Davidson...Allele Biotechnology that now can be deposited at Addgene. What makes these fluorescent proteins unique and...was then screened through successive rounds of mutagenesis for the brightest teal fluorescent proteins that...

...the RBS and allows translation of the reporter gene resulting in GFP signal. With the addition of standards...a ribosome binding site (RBS) followed by a reporter gene. The hairpin blocks translation by sequestering...study the microbiome by using engineered genetic tools and reporter microbes. Let’s take a look at a few ...well as a selectable marker and a desired genetic payload (ex: GFP). Thus, cells can be both sorted using...antibiotic resistance markers, promoters, and inducible gene expression systems in bee gut microbiome species...or preparation, and deliver CRISPR components for gene disruption or CRISPRi. While designed to study the...address these questions, biologists often use next-generation sequencing. Sequencing the 16S rRNA reveals the...

... SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) to detect single molecules of RNA or...plasmids Tagging human transcription factors with GFP Kevin White’s lab recently deposited a set of CRISPR...plasmids to tag human transcription factors with GFP. As part of the Encyclopedia of DNA Elements (ENCODE...proteins for optogenetic control of CRISPR-Cas9. By combining AcrIIA4, a Streptococcus pyogenes Cas9 inhibitor... a few of the new CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR...adapted for detecting SNPs. Learn more from this Addgene blog post Read the paper in Science Find the...the Cas14 plasmids Anti-CRISPR proteins for optogenetic control of CRISPR-Cas9 Anti-CRISPR proteins that...

...NIR FPs can be used similar to GFP-like probes by supplying a single gene to cells. Still in bacteria, ...molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can tell us about cellular events...brightness than previously reported widely used dimeric iRFPs (Table 1). In general, the monomeric state of...normalization to fluorescence of co-transfected EGFP. e Originally reported as a monomer16, IFP2.0 was later found..., which do not produce BV, an additional gene coding for heme oxygenase should be supplied (Baloban et...crosstalk-free three-color imaging with green and red GFP-like FPs (Shcherbakova et al., 2016), as well as ...protein-protein interactions, we recommend using NIR split reporters based on miRFPs (Shcherbakova et al., 2016), since...