...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...

...expression levels compared to AAV9. You can now target these cells with AAV-CAG-GFP packaged in AAV9-X1.1...viral prep pAAV-CAG-GFP AAV9-X1.1 Controls Edward Boyden New serotype pAAV-hSyn-EGFP AAV PHP.eB Controls...Controls Jonathan Ting New viral prep pAAV-CaMKIIa-EGFP AAV1 Controls Bryan Roth New serotype pAAV-Ef1a-DIO...

...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...

...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... to identify why this protein was blue-shifted compared to its parent protein CFP484. Collaborating with...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...

...Fluorescent Protein (GFP) because of the nature of the chemical reaction compared to using a high-intensity...high-intensity laser to rapidly excite GFP. As a result of the different mechanisms to produce photons, chemiluminescent...not required to initiate the reaction. Table 1 compares some general advantages and disadvantages of luminescence... Depending on the specific luciferase used, a compatible substrate must be provided for the enzyme to ...

...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...constructed a library of eGFP-tagged human ORF clones to allow testing and comparison of different cell-free...now available. However, it can be difficult to compare their performance, as most translation initiation...SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of...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...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...

...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... (Sandomenico et al., 2020). Figure 1: Comparison between the IgG antibody and scFv.   What...their lack of an Fc region. Disadvantages scFVs:  Compared to antibodies, scFvs tend to have lower affinities...

... constitutive promoter upstream of GFP increased expression of GFP six-fold, but a stabilized promoter... resulting in variable gene expression that can impact observed phenotypes. Factors such as the growth...medium, growth temperature, and growth rate can all impact plasmid copy number in a cell. One can attempt... origin of replication will be overrepresented compared to those further away. As a result, variable gene...located near the origin will be over-represented compared to those further away. While constitutive promoters...gene expression during shifts in growth rate as compared to a constitutive promoter. To simulate the increase... from a plasmid to the genome without greatly impacting the level of gene expression. For example, the...

...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... variations in DR sequences. Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes when... of performing similarly to Cas9 screens in a compacted library. Overall, Cas12a’s ability to target multiple...or Cas12a. Despite its small size, CasMINI is comparable in efficacy to dCas12a systems in both gene activation...

...fluorescent voltage indicators, however, fall short compared to GFP-based sensors in terms of response and brightness...levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo and achieved... was also between 1013 - 1014 vg/kg, a dosage comparable to those currently used in human gene therapy...precision of gene expression and reduces toxicity as compared to previous Cre/lox or drug-induced Cre systems...

...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...validated their platform on clinical stool samples by comparison to qPCR and demonstrated that it can also detect...bacteriophage shell protein or a bacterial microcompartment protein. The expression of this fusion is ...

...hiPSCs  EMX1  67% Mouse BM cells from GFP transgenic mouse  GFP  ~60-65% Human hepatocytes  Myd88  ~... primary cells. Comparing Nanoblades to Existing CRISPR Delivery Methods When compared head-to-head with...their editing efficiency and off-target effects compare to existing technologies, and their use for HDR...Nanoblades also had lower off-target effects when compared to DNA transfection of gRNA and Cas9 expression...of RNPs generally having lower off-target edits compared to DNA transfection.  Homology directed repair...both technically challenging and can negatively impact zygote viability. Nanoblades, however, avoid this...editing cargo to difficult to target cells. When compared to existing Cas9 delivery methods, Nanoblades ...

...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.   ... from these studies is that the tag protein can impact the expression of the transgene of interest. In... but not necessarily at the same subcellular compartment as the opsin or chemogenetic receptor. This may...

... 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...and Dymecki, SM. Talking about a Revolution: The Impact of Site-Specific Recombinases on Genetic Analyses...

...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...clusters without accumulation in subcellular compartments such as nuclear speckles. Furthermore, CRY2clust...a 1.2-fold greater extinction coefficient when compared to sREACH. Find the FLIM-FRET acceptors here. ...

...this method through monitoring GFP degradation, this protein compartment was used to optimize D-glucaric... hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun Luo’s lab expresses membrane tethered GFP to enhance...a single enzyme. Compared with previous research, which required protein compartments to co-localize multiple...targeted synaptophysin tagged eGFP. AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE from Hongkui Zeng's lab... antibodies for aminoacyl tRNA synthetases Nanocompartments for yeast bioproduction pathways The CRISPR...Research Center developed a pooled plasmid library encompassing all possible amino acid mutations of the receptor...mutations which do not interfere with ACE2 binding, but impact recognition by specific anti-SARS-CoV-2 antibodies...

...extent they overlap. Figure 4 compares the spectra of enhanced GFP (eGFP) and FITC, the latter being a...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...results in a higher-wavelength emission spectrum compared to APC. This happens due to a process called Fluorescence...

... FRET tools, double dimerization GFPs, and circular permutation GFPs. Hopefully we will have new tools...few new ones that are not used so much but have comparable expression levels to things people do use a lot...committees, and how the culture of the university impacts the number of women reaching senior positions. ... those amazing students that want to have a big impact on the world and tell them to work in plant science...