...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 ...

...pPS808 - N-terminal GFP for yeast expression pET GFP - C-terminal GFP for...gene expression GFP Varies pBABE GFP - Retroviral gene expression pMKO.1 GFP - Retroviral ...C elegans expression GFP Localization pcDNA3-EGFP - C-terminal GFP for mammalian expression... from the SP6 promoter pCS2 mt-GFP - Create myc-tag, GFP-tag fusions for transcription ...expression pcDNA3 GFP LIC cloning vector - C-terminal GFP for mammalian expression...pPD95_75 - C-terminal GFP for C elegans expression Other Fluorescent Proteins Localization...expression MSCV-IRES-GFP - Mammalian retroviral gene expression with GFP selectable ...

... 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...

...pAAV-CAG-FLEX-Archon1-KGC-EGFP-ER2 AAV production plasmid encoding for Archon1 fluorescent voltage reporter, Cre-dependent...Fishell 83895-AAV1 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV2 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV8 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV9 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAVrg pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...element Gordon Fishell 83900-AAV1 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...

...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 ...

...Multiple Multi-luciferase reporter vector including transcriptional reporters for NF-kb, TGF-b, c-Myc, ...Root 105533 pAAV.CMV.Luc.IRES.EGFP.SV40 Firefly CMV AAV expression of firefly luciferase and GFP James Wilson...transcriptional regulation and bioluminescent reporters. Plasmid...Luciferase Plasmid Collection You may also like... Fluorescent Proteins Blog: Luciferase 101 Blog: Technologies...Highlights Empty Backbones Expression Constructs Reporter Constructs Luciferase is the enzyme responsible... common choice in assays ranging from use as a reporter gene in vitro and in vivo to use in biosensors...-lives, making them useful as transcriptional reporters. However, they are not secreted, so cells must...

... 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...

...express fluorescent proteins. ID Name Insert Antibiotic Resistance Purpose PI 17446 pLenti CMV GFP Hygro... 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 .... This library expresses EGFP for easy visualization via direct fluorescence. Morris 115644 Morris CellTag.... This library expresses EGFP for easy visualization via direct fluorescence. Morris 115645 Morris CellTag.... This library expresses EGFP for easy visualization via direct fluorescence. Morris Control Lentivirus...Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector, CMV promoter, Hygro Campeau Viral...on each item's material page. Actual titers are reported with each shipment. For more details about our...

...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...

...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase...Cre), together with expression of fluorescent or enzymatic reporters for cellular assays and animal imaging...Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning and...and expressing your gene of interest with GFP expression.  Cloning and expressing RNAi pMKO.1 puro (Plasmid...

...the widely used green fluorescent protein (GFP) gives a bright green fluorescence when exposed to blue ...like chromoproteins, fluorescent proteins, and luminescent proteins. While fluorescent proteins and luminescence...from fluorescent proteins? Before going into the difference between chromoproteins and fluorescent proteins...light, whereas chromoproteins do not fluoresce at all or fluoresce at very low levels. Chromoproteins in...The ocean is full of striking visuals from fluorescent jellyfish, bioluminescent phytoplankton, and colorful... chromoproteins are actually a subset of the fluorescent protein family. What sets them apart is the way... in nature Compared to the history of fluorescent proteins in molecular biology research, chromoproteins...

... Posts on Fluorescent Proteins Browse Our Fluorescent Protein Pages Find the mOX Fluorescent Proteins ...protein non-fluorescent. Unfortunately, accumulated misfolded and nonfunctional (dark) fluorescent proteins...Costantini, and Erik L. Snapp. "Superfolder GFP is fluorescent in oxidizing environments when targeted via...applicable to many microscopy experiments with fluorescent proteins (FPs) localized to compartments other...that they have missed the impact of dark, non-fluorescent, and misfolded FP-fusions on quantitative imaging... and cell physiology in general. Pitfalls of Fluorescent Protein Fusions Observing a bright signal for... fail to correctly fold? Misfolded FPs do not fluoresce. A dark population is not readily apparent and...

... 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...

...: Serotype Testing AAV These AAV encode fluorescent reporters and can be used to compare the tropism of...AAV1). AAV Vectors for Serotype Testing pAAV-CAG-GFP (Plasmid #37825) Description : Ready-to-use AAV in...plasmid 37825 (deposited by Edward Boyden ) and direct GFP expression from the CAG promoter. For information... available from Addgene's viral service. Control EGFP vectors in various serotypes for serotype testing... 37825-AAVrg.T 20 µL $ 140 Add to Cart pAAV-hSyn-EGFP (Plasmid #50465) Description : Ready-to-use AAV ...plasmid 50465 (deposited by Bryan Roth ) and direct EGFP expression from the human synpasin promoter. For...

...neomycin, puromycin, hygromycin) and fluorescent protein reporters (GFP, BFP, RFP, no FP). The vectors each...region of the RPL41 gene, to drive the selection/reporter parts of each construct. This new tool kit allows...

...popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following...in-frame fluorescent fusion protein will allow you to visualize YGOI in vivo (try pcDNA3-EGFP). If you...animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g...

...small molecule activates the fluorescence of the small molecule. Fluorescent RNA aptamers are used to track... applications of fluorescent RNA aptamers in Addgene’s collection. Green fluorescent aptamers Spinach ...selection of an RNA mimic of green fluorescent protein by fluorescence-based selection and directed evolution...using aptamers might know this best, as most fluorescent aptamers are named for a fruit or vegetable! ...What is an RNA aptamer? RNA is not inherently fluorescent and can be challenging to track intracellularly... affinity to small molecules to induce their fluorescence. Until a binding event occurs, neither the aptamer... aptamer nor the small molecule are strongly fluorescent but the binding of an aptamer to its target small...

...sequence of interest and an inactivated fluorescent RNA reporter. If the target sequence is present in ... the RNA reporter will be cleaved resulting in activation of the fluorophore. The fluorescent signal can... a superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage...programmable RNA-binding protein, creating the fluorescent fusion protein dLwaCas13a-NF for in vivo RNA ...

...ACE) in real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry...30220597. ACE system: A versatile in vivo fluorescent reporter for rapid quantification of base-editing...Harris lab has deposited a reporter plasmid that allows for rapid fluorescence-based quantification of base...ratio of mCherry-positive to eGFP positive cells through fluorescence microscopy or flow cytometry. Using...cobalamin-binding riboswitches that bind to cobalamin-fused fluorescent dyes; binding of the cobalamin fusion to the ...the riboswitch results in increased fluorescent signal. Prior to the development of these probes, fluorophore...fluorophore binding aptamers and MS2-binding fluorescent proteins were the state of the art in RNA imaging...

...Plasmid 68295 GFP E17TAG Plasmid 68296 GFP S2TAG Plasmid 68297 GFP Q157TAG Plasmid 68298 GFP S2TAG/E17TAG...expression vector encoding a split mCherry fluorescent reporter, enabling the detection of phosphorylation-dependent...phosphosite, mCherry fluorescence is restored, so interactions can be discovered by fluorescence-activated cell...E17TAG Plasmid 68299 GFP E17TAG/Q157TAG Plasmid 53225 MBP-MEK1 S218TAG/S222TAG ( aka. PCRT7 tetR pLtetO MBP-MEK1...68305 Beta lactamase S68TAG Plasmid 69118 E17TAG GFP zeo resistance Strain 34929 BL21ΔserB Strain 52055...

...such as drug resistance (Chen et al., 2013), fluorescence (GFP; Tzur et al., 2013), or rescue of mutant phenotypes...epitopes, such as GFP (Paix et al., 2014). A further development of this approach reported in 2017 uses the...approaches may become even more powerful following a report that one can use linear repair templates (ie. PCR-derived...templates (Farboud et al., 2019).   The Mello lab also reported that modifying the 5’ modification of oligos used...efficiencies, obtaining as many as 100 independent GFP knock-ins from a single injected animal.  PAM sequence...mixtures (Doksin et al., 2018). The Mello lab also reported efficient editing using a co-injection marker ...

... which makes them attractive for imaging and fluorescence microscopy experiments. The C. elegans genome...genome is well studied and hundreds of reporters have been developed and characterized which allow researchers...This includes CRISPR, RNAi, empty backbones, fluorescent proteins, and Cre expression plasmids. Large ...vectors for C. elegans research, including lacZ and GFP fusion vectors. C. elegans optimized fluorophores...Approach to Better CRISPR/Cas9 Editing Efficiency Fluorescent Tagging of Endogenous Genes with SapTrap Even...

...phosphatase-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked mCherry...cyan-excitable red fluorescent protein (mCyRFP1). Derived from cyan-excitable organge fluorescent protein (CyOFP1...as a donor in FRET (fluorescence resonance energy transfer) and FLIM (Fluorescence Lifetime Imaging Microscopy...elements and combined it with a variety of reporters (i.e. eGFP) and effectors (i.e. DREADDs). The team ...are available at Addgene. Kim, et al. Stem Cell Reports. 2015. PubMed PMID: 25772473 Targeting and manipulating...Segment on This New AAV Tool    mCyRFP1: Novel fluorescent protein that enables imaging of two biosensors...to that of CyOFP1, a large stokes shift, long fluorescence lifetime, and a stable monomeric form as determined...

...low levels of both fluorescence and sequence overlap (coral mOrange2, jellyfish EGFP, and sea anemone mKate2...cells. The Brainbow-1.0 construct contains three fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (...recombination. Brainbow-1.1 scales up the system to four fluorescent proteins, with recombination events specified...but the principles remain the same. The first fluorescent protein in the construct, OFP (orange), is expressed...-2.1 construct can express one of four colors (n-GFP, RFP, YFP or M-CFP.) The construct contains two tandem... tandem invertible segments, each with two fluorescent protein coding sequences in opposite orientations...expressed; once Cre is removed from the system, the fluorescent protein whose coding sequence is closest to the...

...27. See More Fluorescent Proteins Find information on Addgene's collections of fluorescent proteins, including...Preps iPSC Differentiation Factors Antibodies Fluorescent Proteins Other Resources Neurodegenerative Disorders... S A. 2023 Aug 22. Use a lentiviral FRET-based reporter to study tau seeding in mammalian cells. Lathuiliere...Cre-dependent expression of diphtheria toxin receptor (DTR)–GFP fusion protein. Azim et al. Nature. 2014 Apr 17. ... (AIS) plasticity with a motor neuron-specific reporter and a PAX7 inducible vector . Harley et al. Cell...

...for genome editing, empty backbones, fluorescent proteins, reporters, biosensors, and Cre expression plasmids...markers, somatic mutations, or labeling with fluorescent proteins to visualize and track clones of cells... Albert Pan and Alex Schier Labs. Multicolor fluorescent labeling in zebrafish Whole organism lineage ...studying diseases, are listed below: Lipid droplet reporters - Richard White Lab. An imaging platform to study...study lipid dynamics in vivo. Bimolecular fluorescence complementation (BiFC) tools - Marco Morsch Lab...Michel Bagnat lab. Targeting cassettes encoding fluorescent proteins for tagging genes at endogenous genomic...using red light. LipoGlo - Steven Farber Lab. A reporter system that allows direct assessment of lipoprotein...

...designed to test the endocytosis of a fluorescent dye and transduction of GFP-expressing AAV particles we did...following sucrose treatment. Moreover, several reports have shown that cells treated with sucrose or other...

...quantity inside a cell Plasmids 101: Green Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor ...expression plasmids, gene knock-down plasmids, reporter plasmids, viral plasmids, and genome engineering...

...evaluation of shRNA efficacy using fluorescence; use with pGFPns-reporter for cDNA target rtTA On Gu 60495...option rtTA On Kowarz 16623 pBI-GFP Expression of your gene of interest & GFP from a bidirectional tet-responsive...Either Lung 11662 pPRIME-TET-GFP-FF3 Lentiviral, miRNA expression (PRIME) system for application in knockdown...cells; Expresses firefly luciferase hairpin and GFP under pTREtight promoter None Either Elledge 35625...Safe Harbor Locus. Tet-On 3G On Doyon 58245 pGLTR-X-GFP Single vector lentiviral Gateway RNAi system for ...generation; contains expression cassette for TetR-P2A-GFP; see article for additional constructs TetR On Geley...inducible promoter at its 3' end, which controls GFP expression tTA Off Verma 26803 pEnt L1L3 EF1a-tTA...

...there is no fluorescence background resulting from nonspecific donor labeling. Fluorescent proteins (FPs...incorporation of fluorescent probes into protein: hexahistidine-tag-mediated fluorescent labeling with (...Experiments Beware of Fluorescent Protein Aggregation Brush up on Your Fluorescent Protein Basics Resources...Browse Empty Plasmids to Fluorescent Protein Fusions Check out Our Fluorescent Protein Guide ... permeabilized cells  Cy5NTAc 32 25-45 GFP (fluorescein is similar)   Lumio Red Tc: CCPGCC 48 38-70...answers. What is FRET and what's it good for? Fluorescence resonance energy transfer (FRET) is a biophysical...uncoupled donors. The best method for targeting a fluorescent donor to your protein is to hardwire the molecule...

...reproduced their EGFP findings since the initial report in 2023.)     Figure 1: Fluorescence intensity...a magnetoresponsive fluorescent protein. Controlling an optogenetic or fluorescent tool with magnets would... do fluorescent proteins respond to magnetic fields?” and “What are magnetoresponsive fluorescent proteins... expressing good ol’ EGFP — showed a slight but detectable change in fluorescence when a small magnet ...see if any other fluorescent proteins display a similar effect (many do). Fusing EGFP to the self-labeling...set out to do just that — by developing a new fluorescent protein tool that responds strongly to magnetic...Motivated by curiosity Although a wide range of fluorescent proteins and optogenetic tools exist, humans ...

...along with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to ...at equivalent levels upstream of an IRES fluorescent reporter for easy detection.5  Just released: Addgene's... 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...co-express your gene of interest along with a fluorescent protein or selectable marker, it is easiest to...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...

...with Rosella, a pH-sensitive fluorescent biosensor A pH-sensitive fluorescent biosensor-based assay system..., Hess et al. successfully evolved wild type GFP to EGFP using CRISPR-X and subsequent FACS sorting. They...-ome using a quick, affordable, and ubiquitous reporter assay. In their recent publication, Kroeze et ...with unique tissue-specific promoters expressing GFP, tetracycline response elements, and shRNAs many ... of DsRed connected to a pH-sensitive variant of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The...DsRed is relatively pH-insensitive, while SEP fluoresces at cellular pH (7.0) but not at vacuolar pH (...environments that inactivate SEP and leave only DsRed to fluoresce (1). With variants that can be targeted to specific...

...approach to selecting a bright fluorescent protein and a photostable fluorescent protein. In the third post.... In corals, the red fluorescent proteins form tetramers. In general, fluorescent proteins have a natural...your research. But be aware that the fluorescent proteins reported in publications are only characterized... Addgene Blog Which Fluorescent Protein Should I Use? Choosing Your Fluorescent Proteins for Multicolor... with Fluorescent Tags Find Plasmids for Subcellular Localization Browse the entire Fluorescent Protein...select a non-aggregating fluorescent protein. In the jellyfish Aequorea victoria, AvGFP forms a homodimer. ...be compared with mEGFP or mTurquoise2, which are established true monomeric fluorescent proteins. Concluding...

...relies on the fluorescence ratio between the 2 fluorophores, rather than on the fluorescence intensity, ...fluorophore (circularly permuted green fluorescent protein or cpGFP) to the petunia SL enzymatic receptor...the receptor which modulates the fluorescence intensity of the cpGFP without altering that of the internal...straightforward workflow for doing gene targeting, reporter generation, controllable gene knockouts, and inducible...thoughtfully designed plasmid components with Fluorescence Activated Cell Sorting (FACS), allowing for .../doi.org/10.1101/2020.05.04.076067    Novel fluorescent enzyme biosensors provide direct, specific and... Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating a SL-insensitive fluorophore...

...sigh. The problem is that many of these studies report correlations without clearly testing whether a ...Addgene has several great options: pLV hUbC-dCas9-T2A-GFP for removable dCas9 expression and pLM-CMV-R-Cre....expressing these constructs can be achieved by fluorescence-activated cell sorting, and stable gRNA expression...

...explains how to critically assess the reported properties of fluorescent proteins, how to do a head-to-head...comparison of fluorescent proteins and how to make a well-informed decision on the best fluorescent protein ...best fluorescent protein for a specific application can seem like a daunting task. Many fluorescent proteins... number of fluorescent proteins is steadily increasing. Often, the choice for a fluorescent protein is...properties of fluorescent proteins on websites. To make the best choice, the fluorescent protein properties...Brightness Brightness refers to the fluorescence intensity of a fluorescent molecule. A higher brightness increases...tables with fluorescent protein properties and provides a quick way to compare fluorescent proteins, e.g...

... general cloning plasmid, and a prebuilt Unc-32::GFP targeting vector. Jorgensen Lab SapTrap CRISPR/Cas...with the following article: Natsume, et al. Cell Reports 2016 Förstemann Drosophila Cell Tagging System ...provide PCR templates for amplification of the tag (eg GFP, Flag, YFP, etc) and selection markers. Two independent... the first publicly available collection of fluorescently tagged, human, induced pluripotent stem cell...PITCh system plasmids for CRISPR-based knock-in of EGFP-2A-PuroR cassette to the C-terminus of endogenous...the donor vector. The PITCh donor plasmid with an EGFP-2A-PuroR cassette, flanked by microhomologous sequences...first deposited PITCh plasmids were tested by fusing EGFP-2A-PuroR cassette to a nucleolar protein, fibrillarin...

...virus. It also lists several luciferase and fluorescent reporter plasmids that have been used for measuring...libraries . FlipGFP-based activity reporter of SARS-CoV-2 main protease Mpro. (Unpublished) A FlipGFP-based ...method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2). The Broad... termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR). Mammoth Biosciences has published information...FlipGFP-based activity reporter of SARS-CoV-2 protease Mpro now available from Xiaokun Shu's lab. A rapid, highly ...(Link opens in a new window) CRISPR tools and reporters now available from Stanley Qi's lab. COVID-19 ...

...incubation, you assess the level of fluorescence in your cells under a fluorescence microscope. The higher the ...green fluorescent protein (EGFP) reconstitution. (a) Scheme of validation for DSB mediated EGFP expression...where your target is inserted, disrupting the EGFP fluorescent signal, for details on this plasmid, see Mashiko...containing sgRNA sequences into HEK293T cells. The fluorescence intensity was classified into four groups (4...darker than control, 1; very dark). Brighter fluorescence indicates a more successful gRNA design. Adapted...the amount of fluorescence, the better that gRNA is at causing a double stranded break at the target site...screening as little as 8-10 pups. The level of fluorescence you saw in the validation assay should give ...

...pAAV-Syn-Archon1-KGC-GFP-ER2 Syn Archon1 EGFP Constitutive 8 Boyden 115893 pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2] Syn...9 Li Voltage Reporter: Archon 108422 pAAV-CAG-FLEX-Archon1-KGC-EGFP-ER2 CAG Archon1 EGFP Cre dependent...Syn Archon1 EGFP Cre dependent 8 Boyden Voltage Reporter: Voltron 119036 pAAV-hsyn-flex-Voltron-ST Syn...Service: Biosensor AAV Browse our AAV that express fluorescent biosensor proteins, including calcium sensors...Voltron2-ST none Cre dependent 1 GENIE Voltage Reporter: JEDI-2P 179459 pAAV-EF1a-DIO-JEDI-2P-Kv-WPRE ...HaloCaMP1a 138327 pAAV-synapsin-HaloCaMP1a-EGFP Syn HaloCaMP1a EGFP Constitutive 1, 9 Schreiter Calcium Sensor...HaloCaMP1b 138328 pAAV-synapsin-HaloCaMP1b-EGFP Syn HaloCaMP1b EGFP Constitutive 1, 9 Schreiter Dopamine Sensors...

... pX459 (Addgene plasmid ID 48139), which include GFP and puromycin as selectable markers, respectively...CRISPR/Cas9 construct (10 μg total). Add 0.5 μg of GFP expression construct. Electroporate cells with 250...transfection conditions for each cell line with a reporter construct to ensure robust plasmid delivery before...editing efficiency, but 37 °C is acceptable. Fluorescence Activated Cell Sorting (FACS) of Transfected...filter into a FACS tube. FACS sort the top ~3% of GFP positive cells in order to enrich for cells that ...