...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

... 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 ...

...express first with a control reporter gene – GFP for E. coli or mCherry-GFP for mammalian cells. You should...-UAG-sfGFP all TAG sites changes to UAG, RF1 function removed, with Ubiquitin-UAG-sfGFP reporter George...inactivated, with Ubiquitin-UAG-sfGFP reporter George Church 174513 Syn61 No TCG, TCA, or TAG codons in ...TAG Peter Schultz 48696 pANAP AnapRS E. coli fluorescent AA, Anap Mammalian TAG Peter Schultz 49086 pDULE-ABK...Mammalian TAG Peter Schultz 50831 pAcBac2.tR4-OMeYRS/GFP* tyrosyl-tRNA synthetase E. coli various unnatural...or DiZHSeC Mammalian TAG P. Chen 92047 pCOTS-pyl-GFP(35TAG) PylRS M. mazei Cyanobacterial TAG Lital Alfonta...analogs Mammalian TAG Huiwang Ai 160041 pRaGE Pyl TAG GFP Y35TAG PylRS M. mazei Bacterial TAG Lital Alfonta...

...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...