...are generated by inserting a circularly-permuted GFP into an intracellular loop of various G-protein-coupled...enhanced brightness, expression, and sensitivity compared with earlier SF-iGluSnFR. Image from Aggarwal ...size and reduced sensitivity to pH and calcium compared to previous lactate biosensors, and its affinity...

...Church lab uses this system to control expression of GFP.)  To turn this inducible system into a repressible... cells and ~24,000-fold in human HeLa cells as compared to basal expression. Co-expression of repressor...

...labeling. Size is also a factor. Bulkier tags, such as GFP, may interfere with protein expression or function...realizing it. Yet, this selection is crucial as it will impact downstream outcomes of an experiment, including...increased versatility in probe attachment, and compatibility with far-red and near-infrared fluorophores....Grimm & Lavis, 2022). These variations directly impact the way you can utilize these dyes. The most-used...may prove challenging to stain intracellular compartments in in vivo model systems, so extra care must...emission of labeled agonists and antagonists, impacting the interpretation of experimental results. It's...dopamine D2/D3 receptors (Allikalt et al., 2020) Impact of labeling process: The process of dye labeling...

...cassette comprised of OCT4, SOX2, LIN28, NANOG, and a GFP reporter. Minicircles are removed from cells by dilution...efficiency of reprogramming when compared to other methods, even when compared to integrating viral approaches...heterogeneous iPSC cell lines, which can complicate comparisons made between lines. Incomplete silencing of transgenes...and have a higher efficiency of generating iPSCs compared to other delivery methods. Retrovirus has a cloning...lentivirus has similar rates of reprogramming when compared to MMLV-derived retroviruses, it is not as well...heterogeneity between clones. This can complicate comparisons made between samples, i.e. transgene expression...Non-integrating methods have a smaller genetic footprint compared to integrating approaches. These methods eliminate...

...-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked... Listen to Our Tol2 Gateway-Compatible Toolbox Podcast Segment The impact of KLF4 N-terminal variants ...lead to comparable experimental results. Using PiggyBac transposons, the Woltjen Lab compared different...along with EGFP while retaining an emission spectrum that is easily separable from that of EGFP. This makes...latest hot plasmid updates!     Tol2 Gateway-compatiable toolbox for studying the nervous system The... Cole lab recently deposited the Tol2 Gateway-Compatible Toolobox, which will allow scientists to quickly... neuorodgenerative diseases. The Tol2 Gateway-Compatible Toolbox is based on the original Tol2kit generated...

...biological properties of new fusions can be compared with mEGFP or mTurquoise2, which are established true...fluorescent protein. In the jellyfish Aequorea victoria, AvGFP forms a homodimer. In corals, the red fluorescent...detect weak dimerization tendency, as is present in EGFP. Constantini et al, (2012) developed a cell-based...fluorescent protein is monomeric without the accompanying data to prove it is worthless. The OSER assay...that you will be using. If you do a head-to-head comparison right, it will be valuable information for the... are engineered can be shared to simplify the comparison by others. Together, we can build a valuable ...

...enhancer regions (Mendenhall et al., 2013). By comparing gene activation when enhancers were active or ...increased gene activation at the targeted site compared to other Tet1 delivery systems. Transcriptional...) Vlatka Zoldoš’ lab has deposited pdCas9-DNMT3A-EGFP and pdCas9-DNMT3A-PuroR for targeted cytosine methylation...methylation in mammalian cells. Co-expression markers EGFP and PuroR enable sorting and selection of transduced...Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase...epigenome-editing tools. pLV hUbC-dCas9-MQ1(Q147L)-EGFP is also available for lentiviral transduction. Lysine-specific...

...halves of EGFP to recombine by homology directed repair, and resulting in the expression of EGFP. Using ...ordering the sgRNA is simple and easy. Several companies supply gRNA sequences, such as GenScript, ThermoFisher...order them yourself through whatever DNA supply company you currently use. You can also find many previously...indicates where your target is inserted, disrupting the EGFP fluorescent signal, for details on this plasmid,...

..., and disease. In order to expand their model’s impact, organoid users have recently begun to develop ...circularly permuted green fluorescent protein or cpGFP) to the petunia SL enzymatic receptor DAD2. SLs ...which modulates the fluorescence intensity of the cpGFP without altering that of the internal control (Fig...these novel biosensors could have a major economic impact.   Design and mechanism of action of the...

...photobleaching, and achieve better signal in vivo as compared to imaging with broccoli aptamers or MS2 tags ...real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry fluorescence...involves tracking the ratio of mCherry-positive to eGFP positive cells through fluorescence microscopy or...

...Delivering gRNA and Cas9 using adenoviral vectors As compared to other viral vectors, AdVs can provide a large... pAdSh.PGK.Cas9, pAdSh.U6.gRNAS1, and pAdSh.U6.gRNAGFP. Gonçalves says that advantages of AdVs include...guide RNA constructs, pAdSh.U6.gRNAS1 and pAdSh.U6.gRNAGFP, and also Adeno Cas9 and Adeno EA. Or if you're...

...fluorescent proteins, how to do a head-to-head comparison of fluorescent proteins and how to make a well-informed...your selection on a small-scale head-to-head comparative study under conditions that best mimic the intended... QY (EC*QY, sometimes normalized to the value of EGFP). The higher the number, the higher the theoretical...protein properties and provides a quick way to compare fluorescent proteins, e.g. see Chudakov et al (...is of no use. Therefore, it is more sensible to compare fluorescent proteins based on their practical brightness...promising fluorescent proteins and to determine and compare their practical brightness in a system closely ...application. In figure 2 we provide an example of the comparison of the practical brightness of two orange fluorescent...

...Protein Property Visualization allows for visual comparison of FPs and how they relate to one another. .  ...FPbase Fluorescence Spectra Viewer can be used to compare excitation and emission spectra when available,...relating to that specific protein (for instance: EGFP).  Users can “favorite” certain proteins, or create...you or your lab use frequently) for the sake of comparison or ease of retrieval.  These collection pages...store and represent within-study photobleaching comparisons.  We very much appreciate feedback, so try out...

.... This is cumbersome and also makes it hard to compare results across multiple targets because different...primary cells. Find plasmids for CAPTURE 2.0! When compared to the original CAPTURE, CAPTURE 2.0 had a ~14...sequence targeted in the experiment. Figure 2: Comparing CAPTURE and CAPTURE 2.0.     Original CAPTURE...lentiviruses delivering 1) a BioTAP-tagged dCas9 and an eGFP tag, and 2) gRNAs Number of loci targeted One ...regulation. When chromatin conformation data is compared with changes in gene expression, chromatin accessibility...

... complex secondary structure. Today, multiple companies offer long single-stranded DNA production services...maximum Easi-CRISPR efficiency. Easi-CRISPR is compatible with standard microinjection and electroporation...commonly used cassettes, such as fluorescent proteins (EGFP, mCherry etc.), recombinases (Cre, Flp etc.), and...placed to mediate electroporation. Ohtsuka et al. compared traditional microinjection and i-GONAD knock-in...efficiency using a small ssDNA, and they found comparable efficiencies (52 vs 49%). In contrast, Easi-CRISPR...

...non-viral) reporter gene (e.g. firefly luciferase or eGFP) flanked by the EBOV-specific gene start and gene... genes in this plasmid would be more difficult compared to if each gene were encoded in separate (and ...performed in the absence of some viral proteins (compared to viral infection). Therefore, the roles/functions...PMCID: PMC3586226. 4. Mühlberger, Elke, et al. "Comparison of the transcription and replication strategies...

...Please describe any tags or fusions (e.g. His, FLAG, EGFP, etc) on your insert. Use the drop-down menu to ...Addgene Plasmids The Importance of a Fun Workplace: Company Culture at Addgene Resources on the Addgene Website...

...derived from AAV9, their efficacy is generally compared to the AAV9 serotype or an engineered precursor...to be administered concurrently to minimize the impact of neutralizing antibodies developed in response...doesn’t lead to enhanced transgene expression, compared to AAV9, in BALB/cJ mice but is more effective... by the Gradinaru lab (Deverman et al., 2016). Compared to AAV9, it effectively transduces neurons and...toward astrocytes, but its neuron transduction encompasses more molecularly-defined neuronal subtypes and...astrocytes and neurons, with greatly increased efficacy compared to AAV9 (Hanlon et al., 2019). This capsid was...relatively even distribution of target gene expression (EGFP, in the depositing manuscript) across the brain....