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  1. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...three inactivated eGFP reporters- eGFP L202, eGFP L138, and eGFP L93. The modified eGFPs were placed downstream...comparing cells expressing only eGFP expression to cells expressing eGFP and mCherry. ACE was used to identify... To create the eGFP reporters the Harris lab individually mutated three codons in eGFP to eliminate fluorescence...editisome complexes. The eGFP reporters rely on the correction of point mutations in eGFP that results in the...mutated mCherry and a downstream constitutively active eGFP. To create the inactive mCherry, the lab introduced...fluorescence to monitor APOBEC-Cas-9 mediated editing. eGFP reporters for single base editing by APOBEC-Cas9...previous reporter, the Harris Lab created a panel of eGFP reporters to quantify the on-target DNA editing ...
  2. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant...The Kd of EGFP is 0.11 mM [2]. Following the simple logic outlined above, you might think EGFP is unlikely..., cyan and yellow variants of EGFP equally capable of dimerizing as EGFP. As a result, CFP and YFP dimers...contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite multiple studies...protein or Enhanced Green Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general ...using a tag with a serious drawback. The standard EGFP plasmid that used to be sold by Clontech and is ... in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently...
  3. New and Upcoming Viral Vectors - September 2019

    Type
    Blog Post
    ...our entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors... testing AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This...complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available as 20 ul aliquots...recently released these controls: pAAV-hSyn-DIO-EGFP (50457-AAV5): The Synapsin promoter directs broad...broad, neuronal expression. AAV pCAG-FLEX-EGFP-WPRE (51502-AAV5): The CAG promoter directs ubiquitous expression...interneurons. pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5 and 112677-AAVrg): The EF1a promoter...mCherry in the absence of Cre, and expresses nuclear EGFP in the presence of Cre. Biosensor AAV (calcium ...
  4. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

    Type
    Blog Post
    ...pBACKBONE-Flag-hGene(Q295A)-EGFP  Wildtype version: pBACKBONE-Flag-hGene-EGFP or pBACKBONE-Flag-hGene(wt)-EGFP These simple...first. pBACKBONE-Flag-hGene If there was also an EGFP fused to the C-terminal of your insert you would...would list it after the insert. pBACKBONE-Flag-hGene-EGFP Other considerations  If your insert contains a ...
  5. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...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.... Figure 4 compares the spectra of enhanced GFP (eGFP) and FITC, the latter being a commonly used fluorophore...two compounds are completely different in nature (eGFP being a protein and FITC an organic molecule), their... Figure 4: Excitation and emission spectra for eGFP and FITC. As these fluorophores' spectra virtually...expression) and assess our transduction efficiency (eGFP expression).     The first item in Table 2 is a...
  6. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...fluorescent protein (EGFP) reconstitution. (a) Scheme of validation for DSB mediated EGFP expression cassette...halves of EGFP to recombine by homology directed repair, and resulting in the expression of EGFP. Using ...strand annealing) took place and reconstituted the EGFP expression cassette. (b) pCAG-EGxxFP target plasmid...pCAG-EGxxFP target plasmid contains overlapping 5′ and 3′ EGFP fragments under the ubiquitous CAG promoter. The...can be placed in multi-cloning site (MCS) between EGFP fragments. The pX330 plasmid contains humanized ...indicates where your target is inserted, disrupting the EGFP fluorescent signal, for details on this plasmid,...
  7. New and Upcoming Viral Vectors - December 2019

    Type
    Blog Post
    ...Nuc-flox(mCherry)-EGFP 27056 AAVrg pAAV-Ef1a-DIO EYFP 50457 AAV2 pAAV-hSyn-DIO-EGFP   Biosensor AAV...pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP 50457 AAV2 pAAV-hSyn-DIO-EGFP Recombinases Plasmid Serotype... Plasmid Serotype Name 51502 AAV5 pCAG-FLEX-EGFP-WPRE 114471 AAV1, AAV5 pAAV-Ef1a-fDIO mCherry ...
  8. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...Find FRET plasmids here. Split EGFP: An alternative to FRET, split EGFP has also been used to study protein-protein...Fluorescent Protein Mutations Relative to Wildtype GFP EGFP F64L; S65T EYFP S65G; V68L; S72A; T203Y mYFP...protein-protein interactions. In this case, two portions of EGFP are fused to the proteins of interest, and when ...they come into close proximity, the two halves of EGFP undergo folding, maturation, and fluorescence. Biosensors... egfp...
  9. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    ...protein they tested — E. coli expressing good ol’ EGFP — showed a slight but detectable change in fluorescence...better, multiple other labs have reproduced their EGFP findings since the initial report in 2023.)   ...   Figure 1: Fluorescence intensity of EGFP-FlavinTag in E. coli changes in response to a ~25 mT...proteins display a similar effect (many do). Fusing EGFP to the self-labeling FlavinTag made a reliable one-component... the proteins that had shown magnetic responses: EGFP, mScarlet, and AsLOV2. After several rounds of semi-random... semi-random mutagenesis and screening, the EGFP and mScarlet variants were showing no obvious signs of...
  10. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    ...50465 AAV8, 9 (20 µL size serotype testing) AAV-hSyn-EGFP Biosensor AAV The ability to detect the neurotransmitters...Name 119741  AAV5  AAV SYN flex PSAM4 GlyR IRES EGFP Recombinase AAV Finally, we are excited to add...2, 5, rg (20 µL size serotype testing) pAAV-hSyn-EGFP Recombinases Plasmid Serotype Name 55634...Serotype Name 119472  AAV5  AV SYN PSAM4 GlyR IRES EGFP   Biosensors Plasmid Serotype Name 105322... Plasmid Serotype Name 105530  AAV1 pAAV.CMV.PI.EGFP.WPRE.bGH 37825  AAV1 pAAV-CAG-GFP 59462  AAV2...
  11. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ... to LanYFP and the addition of the enhanced GFP (EGFP)-type termini. mNeonGreen was reported as the brightest...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...
  12. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    ...IRES EGFP Chimeric channels for neuronal manipulation 119742  AAV5  AAV SYN PSAM4 GlyR IRES EGFP Chimeric...*  pAAV-CAG-GFP 50465  AAV8*, AAV9*  pAAV-hSyn-EGFP *Also coming in our new 20 ul trial size! Voltage...
  13. Sequencing Primers

    Type
    Guide
    ...forward primer EGFP-C CATGGTCCTGCTGGAGTTCGTG (BD Biosciences) 3' end of EGFP, forward primer EGFP-N CGTCGCCGTCCAGCTCGACCAG...end of EGFP, reverse primer EXFP-R GTCTTGTAGTTGCCGTCGTC (Golenbock lab) For distinguishing EGFP vs ECFP...
  14. Choosing Your Fluorescent Proteins for Multi-Color Imaging

    Type
    Blog Post
    ...proteins for imaging with this set are mTagBFP2, EGFP or one of the improved GFP variants, mRuby2 or TagRFP-T...mammalian cells, one of the improved folding variants of EGFP like mEmerald or Clover is probably best; mNeonGreen...and have reported brightness measurements. Here, EGFP outperforms the improved folding variants, presumably...
  15. New Viral Vectors - Summer 2024

    Type
    Blog Post
    ...Category Depositor  Notes pAAV_hSyn-SIO-stChrimsonR-EGFP-P2A-PdCO-miniWPRE AAV1, AAV5 Optogenetics Yizhar...service with multiple serotypes pAAV_hSyn-PdCO-EGFP-WPRE AAV1, AAV5 Optogenetics Yizhar New viral service...
  16. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    ...pAAV EF1a Nuc-flox(mCherry)-EGFP Brandon Harvey 50457 AAV2 pAAV-hSyn-DIO-EGFP Bryan Roth 50459 AAV1 pAAV-hSyn-DIO-mCherry...pAAV-hSyn-DIO-mCherry Bryan Roth 50457 AAV1 pAAV-hSyn-DIO-EGFP Bryan Roth 114472 AAVrg pAAV-hSyn-mCherry Karl...
  17. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    ...variant library is monitored by fusing the variants to eGFP followed by an internal ribosomal entry site and...a variant has wild-type levels of abundance, the eGFP will be stable and fluoresce. But if the variant...unstable, it will be degraded, leading to reduced eGFP fluorescence. Cells expressing the library are then...fluorescence-activated cell sorting into four bins based on their eGFP:mCherry ratios. Genomic DNA from each bin are then extracted...
  18. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...) 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...
  19. New Viral Vectors - Fall 2024

    Type
    Blog Post
    ... Controls Edward Boyden New serotype pAAV-hSyn-EGFP AAV PHP.eB Controls Bryan Roth New serotype AiP13044...Controls Jonathan Ting New viral prep pAAV-CaMKIIa-EGFP AAV1 Controls Bryan Roth New serotype pAAV-Ef1a-DIO...
Showing: 1 - 20 of 105 results