... Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance...without relying solely on GFP (Katti et al., 2020). They named this reporter “Gene On” or GO. GO works by...mediated 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... their hypothesis with GFP fluorescence. To do this they generated a silent GFP construct, that contains... of eGFP reporters for single base editing by APOBEC-Cas9 editosome complexes. Scientific Reports 9: ....

...following components: a GFP protein that binds to the coding region of a reporter mRNA via a PP7 coat protein...protein (NLS-PCP-GFP) an RFP protein that binds to the 3’ UTR of the reporter mRNA via an MS2 coat protein...64543) Before translation, both GFP and RFP are bound to the reporter, causing it to appear yellow. The...translation pushes NLS-PCP-GFP off of the mRNA, leaving only a red FP bound to the reporter. Because it contains...translation is necessary for removal of GFP from the TRICK reporter. Delayed translation is known to be important... Drosophila patterning gene oskar. Early in development, the osk-TRICK reporter was double labeled, indicating...correlated with GFP intensity, showing that TRICK is an accurate readout of relative reporter translation....

...GBPs/GFP. The new system, CRE-DOG (Cre Dependent On GFP), is activated by GFP and derivatives GFP and ... of GFP. These systems, known as T-DDOG and Cre-DOG, respectively, repurpose popular GFP reporter lines...to bind GFP. The idea of using GFP as a scaffold suddenly seemed very realistic. Envisioning GFP as a substrate...a UAS-driven luciferase reporter construct. In the absence of GFP, no reporter output was observed in ...electroporated T-DDOGs, GFP, and a reporter construct into the mouse retina. GFP expression robustly activated...activated the reporter gene TdTomato, whose expression was absent without electroporated GFP. They then ...T-DDOGs with two mouse GFP reporter lines; again, TdTomato was seen only in cells with GFP, at a high activation...

...apoptosis reporter, ZipGFP, which utilized the self-assembling split GFP (GFP1-10 and GFP11) to increase...self-assembling split GFP (GFP1-9 and GFP10-11) by flipping a single β-strand of GFP, hence the name FlipGFP. FlipGFP... used genetically encoded fluorescent executioner caspase reporters. Several apoptosis reporters have ...fold compared to other reporters (Zhang et al., 2019). How does FlipGFP work? GFP contains 11 β-strands...transfer (FRET)-based executioner caspase reporters. In these reporters, the FRET pair is linked by a caspase...FlipCherry: Red fluorescence protease reporter To add to the fluorogenic reporter toolkit, the Shu Lab also designed...sensor based on tripartite split-GFP association." Scientific reports 3 (2013): 2854. PubMed PMID: 24092409...

...detectable by two different tags: a reporter and an HA tag. The reporter comes in three varieties: HRP, Spaghetti...independent of the reporter. The only caveat is that HA staining is weaker than reporter staining. Picking...Picking the right reporter for your RANBody While all three reporters stain cultured cells and tissue sections... have 10 HA or 10 MYC tags built into a GFP scaffold. Some GFP nanobodies  stain poorly when tagged with...Monster, and Chicken IgY Domain RANbody Reporters. Reporter Detection Method Uses Pros Cons Antigens...weak signals of fluorescent protein (i.e. GFP, RFP) Detecting GFP reconstitution across synaptic partners...recombinant DNA, they can’t be genetically tagged and instead are conjugated to reporters (i.e. fluorescent dyes...

... to stably integrate a reporter gene that contains a puromycin resistance gene followed by the target ...histone)-GFP reporter into the genome of mammalian cells.  Figure 1: The SRIRACCHA reporter construct...that targets your desired gene (whose sequence is also found in the reporter gene). A control RFP expression...Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter systems provide ... CRISPR reporters in the near future.  SRIRACCHA (stable, but reversible, integrated reporter for assaying...that contains a puromycin resistance gene linked to an in-frame H2B-GFP. This visual read out allows for ...will express RFP while cells containing the reporter gene that was cleaved by Cas9 and repaired via HDR...

...scientists to begin to study GFP and deduce its biochemical properties. Now GFP and its fluorescent derivatives...and scientists employ GFP for a wide number of functions, including: tagging genes for elucidating their...constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a promoter of interest...monitor gene expression from that promoter in a given cell type. This type of transcription reporting was ...type may have the GFP under the control of an additional promoter from that of the gene of interest, or ...expressing GFP from cells that are not. Developmental/transgenic uses: Because of its stability, GFP can be...implantation and generation of transgenic mice. Purification: GFP can be used as a general epitope tag for...

...decrease in red puncta. Find the mRFP-GFP-LC3 plasmid at Addgene! GFP-LC3-RFP-LC3∆G: A fluorescent probe ...flux. mRFP-GFP-LC3 functions as an autophagy bionsensor via the same mechanism as Rosella: GFP is quenched...autophagy biosensor is the GFP-LC3-RFP-LC3ΔG probe. It’s similar to the mRFP-GFP-LC3 Tandem Fluorescent Protein...Autophagic flux is estimated by the GFP/RFP ratio: a decrease in GFP fluorescence relative to RFP fluorescence...the mitochondria-targeted Keima variant at Addgene! mRFP-GFP-LC3 tandem fluorescent protein This autophagy...translation, this fusion protein is cleaved to generate one molecule of GFP-tagged LC3 for every one molecule of...decrease in autophagic flux.   Find the GFP-LC3-RFP-LC3∆G plasmid at Addgene! References Klionsky, Daniel J....

...study the A. victoria gene responsible for encoding GFP, and, in 1992, he reported its sequence. Soon after...laboratory. The discovery of GFP The story of how fluorescent proteins like GFP came to be used as research...their work on GFP. This great honor serves to highlight how useful fluorescent proteins like GFP really are...fluorescent ones, like Green Fluorescent Protein (GFP). These molecules occur naturally in bioluminescent... calcium ions, and the second, which we now call GFP, absorbs that blue light and glows green. In the ...new green glowing protein could be used to measure gene transcription, as well as to track protein localization... Prasher’s collaborator Martin Chalfie expressed GFP in exogenous organisms (E. coli, and later C. elegans...

...remove a stop cassette upstream of an effector/reporter gene. In cells where the stop cassette is removed...https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article contributed...that target calcineurin. Current calcineurin gene reporters are based on the bacterial βgal assay, and ...calcineurin transcriptional reporter consisting of a quickly-degraded destabilized GFP variant fused to a 4X...bottom panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (pink) to indicate... Image from Diessl et al., 2020. Find the GFP reporter calcineurin activity here Diessl et al., Microbial...the sgRNAiBAR library covers every annotated human gene, it enables genome-wide screening with a high MOI...

...sensor was engineered from a GFP variant and a DA receptor such that GFP fluoresces when the receptor ...on a similar strategy: a GFP variant bound to a DA receptor, such that the GFP fluoresces upon DA-receptor...Record DA dynamics of optogenetically activated and inhibited neurons in live mice Report modulation of DA ...behaving mice to gather the following types of data: Report time-course and concentration of released DA in...moving mice to gather the following types of data: Report endogenous DA release from a single dopaminergic...looming stimulus in vivo in transgenic zebrafish Report DA release in live mice over the course of Pavlovian...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...line expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to its target... proximity to the GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The...-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP system reports an efficiency of up...ligase TurboID fused to a conditionally stabilized GFP-nanobody rather than a specific protein of interest... For a one kilobase insertion, the researchers reported an editing error rate of less than 0.3%, both ...Fig. 3: Development of a cleavage-free dCas9-based gene editor using microbial SSAPs. Image from Wang, et...using mStrawberry, localization peptides, and reporters that drive the expression of the fluorescent protein...

...Green Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label of cells...remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant...contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite multiple studies ...in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently...best solution is to use truly monomeric FPs. EGFP and other GFP family members (superfolder, Emerald, CFP...However, if you fuse your protein of interest (POI) to GFP to study the protein's behavior in cells, in solution...noncovalently dimerize. That means that your POI fused to GFP or another fluorescent protein (FP) could be forming...

...to further restrict gene activation to cells expressing both Cre and GFP. The GFP dependent FLPo, Flp-...respectively of a GFP dependent Cre. The two segments come together in the presence of GFP to form a functional...extensive supply of transgenic reporter lines that supply a variety of GFP expression patterns, thus allowing...AAVrg pAAV-hDlx-Flex-GFP-Fishell_6 Gordon Fishell   Optogenetics Optogenetics continues to supply neuroscience...-2 proteins ChR2 for targeted expression DREADDs GFP dependent Cre and FLPo And more!  New lentiviral...inhibition library that inhibits over 18,000 human genes and is used for genome-wide inhibition screening...in new serotypes, as well as a new Cre dependent GFP control plasmid for expression in forebrain GABA-...

...dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and...and GFP shRNA. Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown...be used to generate random mutagenesis libraries or to insert heterologous genes, reporters, or other ...they can successfully create GFP fusion proteins with a variety of genes across the genome. Although this...effectively targets your gene/region of interest is a key step for CRISPR/Cas9 gene editing. Dr. Masahito...Masahito Ikawa has created a GFP reporter plasmid for scientists to validate the efficacy of their gRNAs. The...sequence or gene of interest into the pCoofy vectors, select the appropriate pair of vector and gene primers...

...widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have been used... cassette to the gene of interest. The gene to be expressed is amplified using a gene-specific forward...destination vector expressing a separate reporter gene (EGFP, iRFP, IFP1.4, puromycin, neomycin or luciferase...sgRNA-targeted gene, resulting in increased activation of endogenous transcription of the target gene. The brilliant...of downstream genes. Nihongaki et al describe the targeted activation of endogenous genes and detail the...efficiency and gene expression. The pUCXKT vector ensures that all screened plasmids contain a gene variant*...resistance gene (missing the first two codons). Two stop codons are inserted between the truncated gene and ...

...S65T) in the original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness... There are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made...examined on a case-by-case basis. GFP is sensitive to acid The chromophore of GFP can exist in a protonated and...its acid tolerance is maintained in cells.   GFP is big GFP is a 28 kDa protein that resembles a cylinder...2013; Hilaire et al., 2017). GFP can only be used for tagging proteins GFP fusion proteins are made by ... a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP fluorescence overlaps...necessary to generate the strictly monomeric EGFP variant, mEGFP (Zacharias et al., 2002). GFP needs oxygen...

...process worked well with the common GFP variants EGFP, mGFP4-T, and S65T GFP, and no red color was observed...repurpose GFP is great news for microscopy enthusiasts! Addgene has empty backbones containing GFP variants...you’ve used a GFP derivative to conduct subcellular localization studies or make a reporter construct. Fluorescent...exciting GFP-labeled plant cells at 405 nm instead of 488 nm, Sattarzadeh et al. noticed that GFP had turned...cytosol-targeted GFP. White circles represent the irradiated area. Photoconverted GFP spreads through ...Applications and limitations of GFP photoconversion One concern with photoconverting GFP is the expression level...intensity increases with GFP concentration and saturates around 30 uM. For low-abundance GFP fusions, a photoconversion...

...fundamentally the reporter construct monitors the expression of a nearby gene. The reporter gene serves as a...exon of a gene, and thus they do not disrupt the gene expression of the gene they monitor. Gene traps Gene...the gene of interest in any way. For example, if green fluorescent protein (GFP) was the reporter gene...stimulate the expression of the reporter. The system is made up of a reporter gene flanked by a minimal promoter...a promoter-less reporter which needs to integrate within an exon of a gene to generate a transcriptional...predicted genes have unidentified roles as well (Vanni, et al). Is it true that this many genes are truly...? Traps – gene, enhancer, and promoter – all rely on the random integration of a reporter construct in...

...Fluorecent Proteins in Nano Lanterns Reporter gene assays A reporter gene, such as luciferase, usually serves...since become a gold standard in gene expression analysis and a luciferase gene (one of many available to choose...enhancer regions on gene expression can be determined by detection of the reporter in a specific assay...excite GFP. As a result of the different mechanisms to produce photons, chemiluminescent reporters are generally...bacterial cells. Expression of luciferase as the reporter gene is measured via its activity to produce light..., several studies utilized luciferase as a genetic reporter in plant and mammalian cells. Luciferase assays... is also the most commonly used bioluminescent reporter. This beetle emits a yellow-green light with a...

...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...