...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...

...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...

... 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...

... 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...much the spread of the virus to adjacent areas impacted the experiments we wanted to perform. It required...literature, scientists can use the Data Hub to compare different methods and vectors, and use this data... rat CNS: AAV PHP.EB vector dose–response and comparison to AAV PHP.B. Gene Therapy, 25(5), 392–400. https..., C., & Flotte, T. R. (2015). Stability and compatibility of recombinant adeno-associated virus under ...

...GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato, mDlx-GCaMP6f, hDlx-GiDREADD-dTomato...fluorescent tools for labeling structures and compartments in mammalian cells By Angela Abitua Genetically...are often used to label  specific structures, compartments, or specific localization of biomolecules within...usually much easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed...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...

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

...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...labeling in whole tissue (15).  CLARITY is also compatible with LSFM, a method termed COLM (clarity-optimized...spatiotemporal control over labeling methods as compared to transgenic animals (17).  These viral plasmids...

...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...into a translated protein. As depicted in the accompanying cartoon, they have a multitude of uses including...ProtA IgG-binding domains. TAP has since come to encompass several other tag combinations, though frequently...

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