...coding sequence for a green fluorescent protein (GFP). The success of the A-to-G conversion could be seen...through the success of the change in expression of GFP, for the stop codon should have changed from “TGA...cytometry is used to sort populations by mCherry and GFP expression to check for ABE’s. b) Quantification ...

... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...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...

...capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected...In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study) are captured...SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines ...) 3D model of the obtained structure of the H1.8-GFP-bound nucleosome. Images adapted from Arimura et ...

...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...

...considerably faster, with expression levels comparable to GFP during early embryogenesis (Figure 3). mScarlet-I3...through the middle of each embryo are shown, comparing GFP, wrmScarlet (mScarlet), and the new mScarlet3 and... focal adhesion marker paxillin (visualized with GFP). Reprinted with permission from Zhong et al, 2024...shorter than many popular fluorescence markers like GFP (510 nm), allowing co-imaging. And because bioluminescent...

...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A GFP-Rab7A     ...protein Knock-off, is made up of three components: a GFP protein that binds to the coding region of a reporter...reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) a RFP protein that binds to the 3’ UTR of the reporter...these dCas9s with one of three fluorescent proteins (GFP, BFP and mCherry), the authors were able to visualize...monoclonal antibodies against them, fused them to GFP, and compared them to popularly used epitopes tags...

... from the default GFP (green) to RFP, CFP, or YFP fluorescence. In Brainbow 2.1, GFP is tagged with a ...and activate fluorescence of the downstream OFP, GFP, or RFP. Cells that express the construct but do ...generates cells that randomly express either nuclear GFP, membrane CFP, cytoplasmic YFP, or cytoplasmic RFP...incorporates the highly photostable fluorophores mOrange2, EGFP, and mKate2 (red) to solve the problematically low...

... create a fusion with a reporter protein such as GFP? All of these outcomes can be most effectively achieved...typically used for gRNA. May contain reporter gene (e.g. GFP) to identify and enrich positive cells or a selection...transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells. Likely contains...repair by non-homologous end joining (NHEJ) or HDR. EGFP validation of sgRNAs: Can evaluate component delivery...efficacy by cloning in genome target sequence into EGFP reporter. Target DNA reporter system: Can evaluate...

... PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection by ...Joachim Goedhart shares some strategies for generating GFP fusion proteins. From considering protein size and...

...section. The GFP story has its own Franklin in Doug Prasher, who originally isolated the GFP gene but was...the identification of Green Fluorescent Protein (GFP) and its development into an essential tool in biology...

...Since the discovery of GFP over 50 years ago, the growing spectrum of fluorescent proteins (FPs) has ...repair pathway, a new user-defined DNA sequence (like GFP) can be inserted at the DSB. CRISPR has allowed scientists...toolkit offers C- and N-terminal tagging vectors (eg GFP, Flag, YFP, Strep, TEV-V5) with either blasticidin...NanoLuc), fluorescent proteins (TagGFP2, TagBFP, TagRFP, and T2A-TurboGFP-PEST), and small epitope tags ...library containing several types of fluorescent (EGFP, tagRFP, mCherry) and nonfluorescent (Halo, SNAP...

...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...Controls Jonathan Ting New viral prep pAAV-CaMKIIa-EGFP AAV1 Controls Bryan Roth New serotype pAAV-Ef1a-DIO...

...transcriptional activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive...fluorescent signal in the ticker tape from cFos::GFP-iPAK4 within an hour. Analyzing the ticker tape images...to red at 12 h, and PMA activation induces cFos::GFP-iPAK4 at 3 or 6 h. Scale bar 5 µm. Images adapted...

...constitutively expresses a reporter protein (e.g., GFP or luciferase) that is either co-transfected with...result in Figure 1, an internal control, such as the GFP-expressing Plasmid B, could demonstrate whether the...transfection was not successful due to the absence of GFP fluorescence from the living, viable cells. This ...

...substitutions to LanYFP and the addition of the enhanced GFP (EGFP)-type termini. mNeonGreen was reported as the ... to 3 times brighter than the most commonly used GFPs and YFPs. Its excitation maxima is at 506 nm and...protein. In vitro, mWasabi is 1.6-fold brighter than EGFP and is photostable. It also has narrow excitation...it does not significantly excite at 400 nm while EGFP does. Find mWasabi plasmids here! One of the most...

... way. For example, if green fluorescent protein (GFP) was the reporter gene and green cells were observed...developmental stage you could infer that the gene which GFP was monitoring was being expressed in the same pattern...

...experiment, consider co-transfecting with GFP. This allows you to sort for GFP-positive cells and to enrich for...

...imaging with this set are mTagBFP2, EGFP or one of the improved GFP variants, mRuby2 or TagRFP-T, and an... / em 527 nm) can be imaged together but CFP and GFP (ex 488 nm / em 507 nm) show some crosstalk between...mammalian cells, one of the improved folding variants of EGFP like mEmerald or Clover is probably best; mNeonGreen...and have reported brightness measurements. Here, EGFP outperforms the improved folding variants, presumably...

...such as excitation of Green Fluorescent Protein (GFP) because of the nature of the chemical reaction compared...to using a high-intensity laser to rapidly excite GFP. As a result of the different mechanisms to produce...

...binding scFv that’s fused to GFP. When the multiple copies of the scFv-GFP fusion bind the SunTag scaffold...scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based...

...various combinations of protein tags (His, Avi, and GFP) as well as protease sites to facilitate protein ...’ end of the leader sequence. Using a variety of GFP and RFP fusions as their test genes, the researchers...terminators of varying strengths centered within GFP/RFP bicistronic reporters characterized in Cambray...et al 2015). He has also constructed a library of eGFP-tagged human ORF clones to allow testing and comparison...be conveniently performed through the N-terminal eGFP tag carried on all constructs. The Cell-Free Protein...

...each recruits a different fluorescent protein (BFP, GFP, or RFP) to the genomic loci of interest. Pairs of...hairpin sequence (sgRNA-Sirius-8XPP7) to recruit GFP to the loci of interest. To determine how the sgRNA-Sirius...

...regulation. To create iGluSnFR, the Looger lab wedged GFP between two halves of a bacterially derived periplasmic...increase in fluorescence intensity from the inserted GFP. After characterizing iGluSnFR in vitro, showing ...generation of iGluSnFR, the Looger lab swapped eGFP for sfGFP to create SF-iGluSnFR. This improved variant...

...Antibody, Addgene gave to me…  A recombinant anti-GFP!    🎶On the third day of Antibody, Addgene gave ...

... constitutive promoter upstream of GFP increased expression of GFP six-fold, but a stabilized promoter...

...selectable marker and a desired genetic payload (ex: GFP). Thus, cells can be both sorted using FACS and isolated...allows translation of the reporter gene resulting in GFP signal. With the addition of standards, the assay...

...levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo and achieved...voltage indicators, however, fall short compared to GFP-based sensors in terms of response and brightness...

...anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling...This set of new anti-FLAG frankebodies includes mEGFP, mRuby2, iRFP713, SNAP-tag, and HaloTag fusions,...fusions, while the anti-HA frankenbodies include mEGFP, mCherry, and HaloTag. Figure 1: (a) Anti-FLAG...

...interest (analyte), and accumulation of ATP activated GFP expression (receptor), then the transducer could ...plate reader, or any other instrument which measured GFP levels and converted that signal into a readable ...

...protein of interest. The green fluorescent protein (GFP) and mCherry, two of the most common tags currently...article on the Addgene blog: Fluorescent Proteins 101: GFP Fusion Proteins – Making the Right Connection.   ...

... should contain a reporter DNA sequence (such as GFP) in the sense orientation, followed by coding sequence...et al. Nat Biotech 2003): In the presence of Cre, GFP expression is lost, allowing for rescued expression...

...plasmids described in this work including the soCoChR-GFP plasmid as well as several derivatives encoding alternative...ShadowG, a novel dark green fluorescent protein (GFP) with an absorption peak of 486 nm and ShadowY, a...

...hiPSCs  EMX1  67% Mouse BM cells from GFP transgenic mouse  GFP  ~60-65% Human hepatocytes  Myd88  ~...

...overlap. Figure 4 compares the spectra of enhanced GFP (eGFP) and FITC, the latter being a commonly used fluorophore...to fuse them with a fluorescent reporter, such as GFP or mCherry, through genetic modification. As your... you can also use fluorescent reporters, such as GFP, and fluorophore-conjugated antibodies together in...vector also encodes for eGFP, which serves as a transduction reporter. Note that eGFP is not fused to CD45...-Cy7. Next comes our transduction reporter, eGFP. As eGFP emits fluorescence by itself and does not need...cytometer which channel to use to detect eGFP. As you saw in Figure 4, eGFP strongly overlaps with FITC, hence...two compounds are completely different in nature (eGFP being a protein and FITC an organic molecule), their...

...drug resistance (Chen et al., 2013), fluorescence (GFP; Tzur et al., 2013), or rescue of mutant phenotypes...homology arms to knock in large epitopes, such as GFP (Paix et al., 2014). A further development of this...efficiencies, obtaining as many as 100 independent GFP knock-ins from a single injected animal.  PAM sequence...

...red light instead of blue to excite a molecule of GFP. Because red light has approximately half the energy...light, two photons of red light are needed to excite GFP compared to only one of blue. Only at the focal point...infinitely small point of light – think of a single GFP molecule – will appear as a fuzzy 200-300 nm spot...

... hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun Luo’s lab expresses membrane tethered GFP to enhance...  After validating this method through monitoring GFP degradation, this protein compartment was used to...targeted synaptophysin tagged eGFP. AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE from Hongkui Zeng's lab...

... FRET tools, double dimerization GFPs, and circular permutation GFPs. Hopefully we will have new tools...

...tMAS::rLUC-I::pMAS. Fluorescent proteins (FP, e.g. GFP) can be used in many ways. Often a FP is fused to.... "Evaluation and comparison of the GUS, LUC and GFP reporter system for gene expression studies in plants...

...GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato, mDlx-GCaMP6f, hDlx-GiDREADD-dTomato...pMAK463 (NBαMouse-IgK, Addgene #140701) pMAK464 (NBαEGFP, Addgene #140702) The pMAK461-64 plasmids were...introduced into the hPSC genome using AAV1 and contain an EGFP reporter. Expression of the dCas9-KRAB activator...

... cassette with an ubiquitous promoter to express GFP in a group of young rats. We found that, in this ...label norepinephrine (NE) expressing neurons and GFP (green) to label neurons transduced by each serotype...

...chemistry! The discovery of green fluorescent protein (GFP) sparked a renaissance in biological imaging. Suddenly...advances lead to an obvious question: In this age of GFP and its ilk, why should cell biologists talk to chemists...

...mouse brains expressing green fluorescent protein (GFP) (6).   The Dodt and Frank Bradke laboratories later...subpopulation of neurons are brightly labeled with GFP (e.g. Thy1-YFP-H mice).  However, we recently developed... nerve afferents labelled with a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical...

...protease protection. Thus far Plasmids 101 has covered GFP and its related fluorescent proteins, which are sometimes... I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent Protein Guide Expression Vectors from...

...antibodies against popular fluorescent proteins (GFP and YFP) and protein tags (c-MYC and HA). We have...

...Figure 5: A) Representative confocal images showing GFP expression from ssAAV5s with hIBA1 or hIBA1a promoters...Quantifications showing high microglia specificity of GFP expression for the indicated ssAAV5s. C) Quantifications...

...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 ...

... state-of-the-art NIR FPs can be used similar to GFP-like probes by supplying a single gene to cells. ...crosstalk-free three-color imaging with green and red GFP-like FPs (Shcherbakova et al., 2016), as well as ...for miRFPnanos, 35 kDa for miRFPs, and 27 kDa for EGFP) and a possibility to be used as internal tags due... normalization to fluorescence of co-transfected EGFP. e Originally reported as a monomer16, IFP2.0 was...

...activity can be monitored by phosphatidic acid probe GFP-PASS. B) Time series of HEK 293T cells expressing...membrane-targeted SuperPLDx30 (magenta) and PA probe GFP-PASS (green). After 30 minutes of blue light illumination...

...ends up being translational. Ex: the discovery of GFP. Chapter 4: Goals of the Center for Regenerative ...