...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...identified a family of GFP proteins in the cephalochordate Branchiostoma floridae, named bfloGFPs. They went on...and demonstrate that they can successfully create GFP fusion proteins with a variety of genes across the...enabled scientists to engineer a wide variety of GFPs with diversity in fluorescence brightness, intensity...Cas9 gene editing. Dr. Masahito Ikawa has created a GFP reporter plasmid for scientists to validate the efficacy...plasmid undergoes HR or SSA to reconstitute functional GFP, and the cells will turn green. Mashiko et al.,...

...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...ligase TurboID fused to a conditionally stabilized GFP-nanobody rather than a specific protein of interest...SSAP system reports an efficiency of up to 20%, comparable to commonly used CRISPR/Cas9 methods for large...error rates overall ranged from 0.3% to 10%, in comparison to the 62-90% error rate seen with Cas9 editors...

...electron microscopy analysis is time consuming. GFP-tagged Atg8p or LC3: ATG8 is part of a group of genes...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 ...Fluorescent Protein Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy... changes when it goes from a more neutral pH compartment, ­­like the cytosol, to the higher pH of the ... to other pathways, it is standard practice to compare degradation rates between samples treated with ...

.... https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article...reporter consisting of a quickly-degraded destabilized GFP variant fused to a 4X CDRE repeat. In addition to... Ca2+ (bottom panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (...death. Image from Diessl et al., 2020. Find the GFP reporter calcineurin activity here Diessl et al.,...plasmids for purification of tTALE-eGFP and tTALE-mCherry. In comparison to intercalating fluorescent dyes...microscopy, they also show that this method is compatible with dead cell staining using propidium iodine...

...effects of GFP fusions to inntags or traditional FLAG, HA, and Myc tags. Impressively, IT6-GFP did not reduce... reduce yeast cell size or viability compared to untagged GFP overexpression, while the rest of the fusions...

...TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing GFP that can be...during optimization experiments. This version of GFP will contain a phosphoserine at amino acid 17 instead...instead of a stop codon, which will allow full length GFP to be translated), and second, through a gel shift...protein. Up to 80% p-Ser incorporation into E17TAG GFP has been achieved with the Rinehart system (6); however...bacterial strain – Addgene #68306 pCRT7 NT TOPO E17TAG GFP plasmid - Addgene #68295 SupD plasmid – Addgene #...negatively charged residue, aspartate or glutamate, imparting the negative charge associated with phosphorylation...enzyme function, protein interaction, etc. by comparison to non-phosphorylated or ‘wild type’ control ...

...unbound GFP-dCas9. With these improvements, labeling efficiency was similar to that of a comparable FISH ...signal to noise ratio of the system, they modified GFP-dCas9 and the gRNA scaffold to enhance complex assembly...sequences, which recruit fluorescent proteins BFP, GFP, and RFP fused to the respective hairpin binding ...Fluorescent Proteins Control Protein Activity with GFP Resources on Asddgene.org Find Fluorescent Protein...with the previously established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that...that they could evaluate the degree of chromatin compaction even for this small distance! To the authors’...

...array plus a GFP tag. Four weeks post-infection, there was strong expression of Cpf1 and GFP in the targeted...4 targets when an array of 4 crRNAs, Cpf1, and a GFP tag were transfected on a single plasmid. This is...~75% of neurons were co-transduced with Cpf1 and GFP.  Of these neurons, ~15% had indels at all 3 targeted...Cas9.  For a brief comparison of Cpf1 vs. Cas9, see the table below. Table 1. Comparing the Cas9 and Cpf1...simplified system for editing multiple targets compared to Cas9.  Read on to learn more about Cpf1 multiplexing...lentiviral transduction One vector expresses Cas9, eGFP, and up to 4 gRNAs Each gRNA requires its own promoter...expression vector.  See the graph in figure 2 for a comparison of editing frequency resulting from transfection...

... to track where GFP labelled B cells were in mounted slices of these organs. These GFP labelled cells ...enabled by fluorescent proteins The optimization of GFP and its derivatives by the late Roger Tsien changed...stimulus. Using an epifluorescence microscope and GFP fusion proteins controlled by promoters specific ...the immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph... the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after infection...the cells mechanically in a vibrating nozzle, imparting a positive or negative charge to cells that fluoresce...Laser include: Blue-green fluorescent proteins like EGFP Yellow-orange fluorescent proteins like TagRFP, ...

...includes a set of plasmids each containing an anti-GFP nanobody fused with a different functional element...using different detection methods. Since the anti-GFP nanobody is able to bind fluorescent proteins derived...here can be used to target any protein fused with GFP, YFP, mCerulean, or other related fluorophores. Find...faithfully transcribes RNA to DNA with low error rates compared to other RTs. MarathonRT represents a major step...

...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...difficult to compare the performance of different nucleases. Nicola Patron’s lab compared the efficiency...more predictable and precise deletion products compared to monomeric Cas9’s. Read the paper in Nature...

...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 ...several pooled libraries: Cas13X and Cas13Y, two compact Cas13 proteins found in the metagenome datasets...

... 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...Purity Purity of AAV preparations is assayed by comparing the relative stoichiometric ratios of the viral...

... Vacuolar Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B ...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A GFP-Rab7A     ...monoclonal antibodies against them, fused them to GFP, and compared them to popularly used epitopes tags such ...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...

... 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...your expression system and delivery methods are compatible with all your components. Next, consider the ...screens using CRISPR. AAV transduction Only compatible with SaCas9 (packaging limit ~4.5kb). CRISPR ...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...

...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...  Here's what you'll find in this post:  New compact, efficient prime editors A versatile and precise...tape Teaching an old antibody new tricks   New compact, efficient prime editors By Susanna Stroik The ...Phage-assisted evolution and protein engineering yield compact, efficient prime editors. Cell, 186(18), 3983–4002...the range of targetable cell types. DIRECTED is compatible with fusogens from different viral families and...

...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 ...needs a standard against which the results can be compared; results from an experiment lacking the proper...baseline/standard against which other samples can be compared. The Empty Vector Control (mentioned above) could...

...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...and sgRNA-Sirius-8XMS2 scaffolds performed, they compared them head-to-head against CRISPRainbow. Both tools...visualization tools can be used in live cells. Compared to its predecessor CRISPRainbow, CRISPR-Sirius...

...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... interested in are compatible with your microscope optics, you will want to compare the excitation and...fluorescent proteins currently available. Choosing compatible fluorescent proteins To choose a set of fluorescent...scope of this post. Are your fluorescent proteins compatible with your microscope optics? To determine if ...is efficiently collected by the microscope. To compare the match between a fluorescent protein and a filter...here's a screenshot from the Chroma Spectra Viewer comparing a standard Cy3 or Rhodamine filter set (Chroma...

...experiment, consider co-transfecting with GFP. This allows you to sort for GFP-positive cells and to enrich for...system, developed by the Zhang lab, which has comparable efficiency to the optimized chimeric design but...