... to use a suite of fluorescent reporters. Bacteria containing unstable fluorescent proteins under the ...to reduced eGFP fluorescence. Cells expressing the library are then sorted by fluorescence-activated cell...inoculation, no more fluorescent proteins are made. As the bacterial replicate, fluorescent proteins will be...colonizer, labelled with a constitutively fluorescent reporter can also be used to track how one population...growth rate using TIMER and an unstable fluorescent protein reporter under the control of promoters that ...Addgene Blog Visit the fluorescent protein topics page Get the basics on GFP Find microbiology blog posts...wild-type levels of abundance, the eGFP will be stable and fluoresce. But if the variant is unstable, it...

...Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells GFP oligomerization can lead to ...Janelia Research Campus reminds us that, although fluorescent proteins are incredibly useful, chemical fluorophores...Lab at MIT developed plasmid based systems that report the order in which specific chemical inputs are...recombinase-based state machines could potentially be used to report upon and/or engineer “temporally ordered biochemical...derived from Zika virus and, upon binding, produce reporter proteins. These toehold switches can be used to...viral replication and transcription as well as reporter genes. To learn how you can use minigenomes in...to different pH’s and some that are fused to fluorescent proteins. She also showed that R-bodies can rupture...

...NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...better biosensors. NanoLuc®-fluorescent protein fusions for imaging Fluorescent proteins are fantastic imaging...more penetrative orange fluorescent protein fusion (OgNLuc). Both fluorescent proteins can be excited ...has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged...for infection of host cells. The fluorescent moiety of the BRET reporter was used to sort successfully infected...set out to design a better fluorescent protein that could be co-imaged with GFP using the same wavelength...cyan, yellow, green, orange and red fluorescence. The paper reports a wide variety of applications of the...

...measurements, specifically fluorescent ones? Many of the available biosensors use a single GFP or CFP/YFP-derived...available. Detection methods Many calcium reporters use a single fluorescent protein (FP), requiring only a simple.... This sensor switches from green fluorescence to a red fluorescent state only by simultaneous (1) user-controlled...Genetically encoded calcium indicators (GECIs) are fluorescent proteins linked to calcium binding proteins (... (e.g., calmodulin or troponin C). Their fluorescent readouts are used to monitor calcium concentrations...GCaMP6 family of sensors are commonly used calcium reporters with different kinetic parameters. There are three... of the sensor, and (3) subsequent change in fluorescence only occurs at high calcium concentrations. ...

... microbes rely heavily on fluorescent protein genes. These optical reporters have limited deep-tissue ... consists of a library of reporter plasmids, each containing two fluorescent proteins and four repeats...collection of reporter plasmids is amenable to Golden Gate assembly for turning the single-input reporters into...and a reporter for tracing the engineered cells in vivo, the non-immunogenic violet-excited GFP variant...Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive imaging of microbes in mammalian...contributed by Lukas Morgan  Listen to the acoustic reporter gene podcast segment In recent years, scientists...Mikhail Shapiro’s lab has developed four acoustic reporter gene (ARG) plasmids to better monitor gene expression...

...time.Hallmarks of fluorescent protein timers When FP timers are first expressed, they predominantly fluoresce one ...independently) originally fluoresce green or blue and, when they mature, fluoresce red. The ratio of the ...., et al. "Monomeric fluorescent timers that change color from blue to red report on cellular trafficking...Addgene Blog Learn More About Fluorescent Protein Aggregation Catch Up On Your GFP Basics Get Advice on Choosing...Choosing Your Fluorescent Protein Resources at Addgene.org Find Fluorescent Protein Timers Browse All...All Fluorescent Protein Empty Backbones Check Out Our Fluorescent Protein Guide ...Even before fluorescent proteins (FPs) came into wide use, there were a variety of ways to monitor cell...

...plasmid that constitutively expresses a reporter protein (e.g., GFP or luciferase) that is either co-transfected...transfection was not successful due to the absence of GFP fluorescence from the living, viable cells. This result... well of your cells. Regardless, the amount of reporter protein activity correlates to both the amount...result in Figure 1, an internal control, such as the GFP-expressing Plasmid B, could demonstrate whether the...successfully and expressing the protein. For example, fluorescence microscopy images resulting from our experiment...produces the expected effect, which is green fluorescent cells (Figure 3).   Figure 3: Expression of Plasmid...

...when paired with appropriate fluorescent optics. Existing red fluorescent voltage indicators, however,...levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo and achieved...31937940. Detection of neural activity using a red fluorescent genetically encoded voltage indicator Article...however, fall short compared to GFP-based sensors in terms of response and brightness. They also do not spectrally...voltage-sensitive inhibitory rhodopsin (Ace2N) with the red fluorescent protein (mScarlet) with either a seven or eight...manipulations when combined with existing green-fluorescent GEVIs and blue-light-sensitive channelrhodopsins...indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed PMID: 31685893. A MoClo toolkit for...

... an ERK kinase translocation reporter with a mRuby2 fluorescent reporter (ERK KTR-mRuby2). The ERK KTR...with GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable fluorescent proteins... the active site. Dronpa also produces green fluorescence when exposed to 500 nm light.  Under 400 nm ...active, it phosphorylates the ERK KTR and red fluorescence localizes to the cytoplasm. When MEK1 and ERK...inactive, the ERK KTR isn’t phosphorylated and red fluorescence can be found in the nucleus. For testing kinase...Addgene Blog Learn More about Photoactivatable Fluorescent Proteins Control Protein Expression with Cre-lox...Photoswitchable Fluorescent Proteins ...

...What are Acoustic Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used... to study gene expression in cells. But these reporters have limited use in vivo because they are based...expression in vivo.  But what ultrasound lacked was reporter genes that could produce ultrasound contrast. ...our lab - the Shapiro Lab - developed Acoustic Reporter Genes (ARGs), which are sets of genes that, when...without signs of toxicity or burden relative to a fluorescent protein control. We also added the toxin-antitoxin...P2A-linked accessory genes gvpNJKFGWV marked with Emerald GFP. These plasmids can be used directly in transient...gating for the top quartile (or higher) of BFP and GFP producing cells. After FACS, return the cells to ...

... blog Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent protein... co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence localizes to the plasma membrane ...2022.08.13.503850.  iGECInano NIR calcium reporter Near-infrared (NIR) fluorescent proteins promise better performance...area with lots of bright new plasmids is our Fluorescent Biosensors Collection — scientists are always...measure! Let’s take a look at some of the hottest fluorescent biosensors new to Addgene this year. Feel the...Hiroshi Hibino and colleagues devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular...region-to-IntrAcellular loop linkers” (MTRIA for short), where a fluorescent protein is inserted into one of the loop regions...

...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...

...third-generation intensiometric glutamate sensing fluorescent reporter (iGluSnFR3) to study glutamate, a signaling...can be transferred to the roGFP2 cysteine thiols and change its fluorescence properties. The biosensor... for awards to early career scientists using fluorescent proteins in their research. These awards help...roles in the development and dissemination of fluorescent protein tools. This year, we’ve made some changes...research.   Abhi Aggarwal: Glutamate sensing reporters Abhi Aggarwal is a postbaccalaureate research ...disease,” says Aggarwal. This biosensor changes fluorescence based on the release of glutamate from neurons...line, Hep3B. From left to right: Mitotracker Red, GFP for the biosensor, Hoechst, and overlay).   ...

... transformation. Choose good reporter genes There are many great reporter genes available and which one...pSB166 - pL2_pSB90_tMAS::rLUC-I::pMAS. Fluorescent proteins (FP, e.g. GFP) can be used in many ways. Often ...Evaluation and comparison of the GUS, LUC and GFP reporter system for gene expression studies in plants...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...

...will not produce the fluorescent protein while areas that are exposed will fluoresce. With these images,...the DNA-binding domain of AraC using a mCherry reporter downstream of a PBAD promoter. While none of the...contains BLADE, the PBAD promoter, and a mCherry reporter downstream of the PBAD promoter. To use this plasmid...strain transformed with pBLADE expressing superfolder GFP instead of mCherry under the PBAD promoter. They ...

... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...expression and/or function. These data are sometimes reported or posted on the material page for the corresponding...viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, ...prep # 55632-AAVrg). pAAV-CAG- FLEX-rc [Jaws-KGC-GFP-ER2] was a gift from Edward Boyden (Addgene viral...recipient’s initial thaw will be accounted for in our reported titers. Lentiviral vectors are titered using a...

...viral prep pAAV-CAG-GFP AAV9-X1.1 Controls Edward Boyden New serotype pAAV-hSyn-EGFP AAV PHP.eB Controls...AAV9. You can now target these cells with AAV-CAG-GFP packaged in AAV9-X1.1!  We also have eight new AAV-PHP.eB...drive expression of either Cre recombinase or a fluorescent marker in the various cell types of the basal...check out Addgene's Data Hub to see viral vector reports from other users. And don't forget to use the link...Controls Jonathan Ting New viral prep pAAV-CaMKIIa-EGFP AAV1 Controls Bryan Roth New serotype pAAV-Ef1a-DIO...

...stability of fluorescence over time is crucial. For instance, the fluorescence of fluorescein isothiocyanate...the b(right)est fluorescent proteinFluorescent Proteins 101: A History of Fluorescent Proteins  ...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes,...dyes, and fluorescent ligands. Figure created with biorender.com. Fluorescence refers to the emission...selection and optimization. Fluorescent experiment types: Different types of fluorescent experiments, such as...to the ability to ‘toggle’ between fluorescent and non-fluorescent states in response to a specific stimuli...technologies in fluorescent dyes/ligands Throughout the history of cell biology, fluorescent labeling techniques...

...Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral sgRNA targeting mCherry reporter (mChe)...marmosets. CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons...colleagues identified a highly bright and stable fluorescent protein, StayGold. Although it quickly saw wide...dimerization while maintaining the bright and stable fluorescence of the original (Ivorra-Molla et al., 2023)....Ivorra-Molla, E., et al. (2023). A monomeric StayGold fluorescent protein. Nature Biotechnology, 1–4. https://doi.org..., et al. (2023). Bright and stable monomeric fluorescent proteins derived from StayGold. ResearchSquare...

...of varying strengths centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013... Using an Orthogonal Ribosome-Controlled Fluorescent Reporter."ACS chemical biology 11.1 (2015): 31-37...various combinations of protein tags (His, Avi, and GFP) as well as protease sites to facilitate protein ...library by in vivo functional screen." Scientific reports 5 (2015). BIOFAB plasmid set 3000 Series Confronted...’ end of the leader sequence. Using a variety of GFP and RFP fusions as their test genes, the researchers...fusions. Some applications of PUF protein fusions reported in the literature include RNA detection, RNA cleavage...et al 2015). He has also constructed a library of eGFP-tagged human ORF clones to allow testing and comparison...