...selection cassette. They could also switch from FLAG to GFP or a different epitope. Addgene: Do you have any ...the function of the protein. Myers: We’ve done comparisons of our new system versus standard ChIP-seq for...

...line, Hep3B. From left to right: Mitotracker Red, GFP for the biosensor, Hoechst, and overlay).   ...(DKFZ) who has developed new redox sensitive GFP2 (roGFP2)-based biosensors for live cells. His work focuses..., Pedre Pérez’s biosensor fuses MPST to roGFP2. Because roGFP2 contains two cysteines near the chromophore... anchoring domain), in electroporated in rat hippocampal cultured neurons, imaged 14 days post-transfection...the MPST hydrosulfide can be transferred to the roGFP2 cysteine thiols and change its fluorescence properties... the screen to understand mouse cortical and hippocampal neural microcircuits. “In the longer term, this...

...Expression with Cre-lox Control Protein Activity with GFP Resources at Addgene.org Find KTR reporter plasmids...signal strength, and how long their signals last all impact transduction of the message. To study signaling...dimeric Dronpa is less prone to aggregation in cells compared to the tetrameric Dronpa it was derived from. ...

... for a researcher to come into contact with than GFP. Similarly, if the viral vector carries an shRNA ...materials, you’ll need to use aerosol-tight caps, compatible with your centrifugation speed, to cover the ...

..., Myc and SV40 Nuclear Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral...variant has similar photostability and brightness compared to the original, though its maturation is even...cell-specific tropism, with slightly reduced efficacy compared to AA9-X1.1. Importantly, AAV1-X1 is not neutralized...

...Lindsey M. Costantini, and Erik L. Snapp. "Superfolder GFP is fluorescent in oxidizing environments when targeted...with fluorescent proteins (FPs) localized to compartments other than the cytoplasm. That is, unfortunately...investigators realize too late that they have missed the impact of dark, non-fluorescent, and misfolded FP-fusions...quantitative imaging experiments or even negatively impact cells (see Figure 1). When behaving properly, ...positioning of the FP sequence relative to cell compartment targeting sequences, which often have absolute...that FPs are often poorly suited for cellular compartments other than the cytoplasm. Generally, FPs evolved.... Many of the resident proteins within these compartments undergo significant post-translational modifications...

...check out our article: Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection. Cellular...expressed transiently by transfection or stably impacts the success of BiFC experiments. In either case...independent of PPIs. Overexpression can be assessed by comparing the expression levels between BiFC constructs ...localization of BiFC constructs should also be compared against those of their endogenous counterparts...association between FPs independent of PPIs. Simply compare the fluorescence intensity of your BiFC constructs...under relatively physiological conditions when compared to FRET. However, it’s important to note that ...

...contrast, the widely used green fluorescent protein (GFP) gives a bright green fluorescence when exposed to...fluoresce at very low levels. Chromoproteins in nature Compared to the history of fluorescent proteins in molecular...eforRed, asPink, spisPink, scOrange, fwYellow, amilGFP, amajLime, cjBlue, meffBlue, aeBlue, amilCP, tsPurple...

... a superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage...the ability to cleave the RNA target.  Table 1: Comparison on Common CRISPR Enzymes Name  Enzymatic Domains...multiplexing; expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although... cell stress, and RNA size distribution. When comparing LwaCas13a to shRNA, they found that LwaCas13a ...

...which must be stabilized by monomeric superfolded GFP and averages only ~50% RNA knockdown. In initial ...off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced off-target sites from the 18,385...edits by ~45% and off-target A-to-I edies by ~94% compared to RESCUE.   Addressing the potential use of RESCUE...

...in this case a reporter gene (e.g. luciferase or GFP), regardless of whether it interacts with a transcriptional...transcription. The HIS3 reporter system (e.g. the Gateway-compatible bait Y2H plasmid pEZY202  from Yu-Zhu Zhang’s...particular cell type, or a subset based on a cellular compartment or growth stage). A prey library can either be...

...In cases such as this, an epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest ... need to choose the protein type that is most compatible with the host species and isotype of their primary...developed, which can be used for most antibodies compatible with either Protein A or G.  While commonly used...

...Mammalian Cells Learn about the Dangers of Using GFP for Protein Localization Resources on Addgene.org...line identity and purity making it easier for comparable studies to be performed on a cell line even decades...through working for a variety of pharmaceutical companies including GSK, AstraZeneca-MedImmune, and Infors...

...SYFP2 Clover Green Mammalian Expresses Clover (a GFP variant) commonly used with mRuby2 pLSSmOrange-...to target mTurquoise2 to various subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian Expresses mammalian...Clover pGWF1 Cyan & Yellow Bacterial Gateway-compatible vector to express a gene of interest fused between...

...used this cassette to knock out genes, knock in a GFP cassette, and introduced site-directed mutations,...populations to retain more of these plasmids as compared to those without the restriction modification ...

...Vectors Read about Reporter Gene like Luciferase and GFP Resources on Addgene.org Find Plasmids for Your...promoter present in your plasmid backbone must to be compatible with the type of RNA that needs to be made: if...

...liver injury markers were not increased compared to a control AAV-GFP. Not only did the AAV-SaCas9-sgRNA constructs...editing is the potential for off-target effects. To compare the off-target effects of SpCas9 and SaCas9, Zhang...had modest editing rates in HEK-293T cells when compared to the full-length SpCas9. In 2016, Chew et al...cleavage of HBB and CCR5 genes in human cells using compact expression cassettes. Sci Rep 5: . https://doi....

...aptamers able to bind to a small molecule derivative of GFP called 3,5-difluoro-4-hydroxybenzylidene imidazolinone...identified through microfluidic-assisted in vitro compartmentalisation for their high binding affinity to TO1-biotin...of these variants exhibit increased brightness compared to Mango, and all are photostable and useful for...

..., Hess et al. successfully evolved wild type GFP to EGFP using CRISPR-X and subsequent FACS sorting. They...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...several GMAP-compatible vectors for a wide variety of biological applications. First, GMAP-compatible backbones...including a stop codon) formats. The collection is compatible with both standard Gateway cloning and the FNLCR...sites. For instance, the authors generated a GMAP-compatible backbone containing Rosa26 homology arms. The...GMAP, there will be a continuous expansion of compatible backbones, promoters, and genes available to ...

...fluorescent protein that could be co-imaged with GFP using the same wavelength for excitation. The result... a good story (1). From a deep sea shrimp to a compact powerhouse of bioluminescence, NLuc is 100-fold...yield, brightness and improved maturation time compared to similar orange fluorescent proteins. In their...more than 2-fold brighter signal from Antares as compared to Fluc in this model. The construct for Antares...transfer of energy from one fluorescent protein to a compatible fluorescent protein. FRET can be measured by ...