...and best solution is to use truly monomeric FPs. EGFP and other GFP family members (superfolder, Emerald...problematic, several reportedly monomeric FPs are not actually monomeric, at least in practical terms. ...FPs, are not necessarily monomeric, even if they were described as monomeric in a publication. The gold....A., et al., Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells. Science...remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant...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...

...reported was PA-GFP. PA-GFP was derived from Jellyfish (Aequorea victoria) GFP (wtGFP) by mutating Threonine...Emission (nm) PA-GFP Monomer UV-Violet Dark/ Green ~200 Medium 504/ 517 PS-CFP2 Monomer UV-Violet Cyan...mutation in PA-GFP generates a chromophore population that is more neutral, causing PA-GFP to display almost... 583/ 593 mKikGR Monomer UV-Violet Green/ Red 560 High 580/ 591 Dendra2 Monomer UV-Violet or blue ... (nm) Dronpa Monomer UV-Violet Dark/Green N/A (high) High 503/ 518 Padron Monomer Blue Dark/ Green...503/ 518 rsCherry Monomer Yellow Dark/ Red 7 Low 572/ 610 rsCherryrev Monomer Blue Dark/ Red 20 Low.... Both variants are more monomeric than mEos2 (mEos4b is completely monomeric) and display bright and ...

...the enhanced GFP (EGFP)-type termini. mNeonGreen was reported as the brightest monomeric green or yellow...plasmids here! mTFP1: a monomeric teal fluorescent protein mTPF1 is a monomeric version of the tetrameric...bright, monomeric green fluorescent protein. In vitro, mWasabi is 1.6-fold brighter than EGFP and is photostable... to 3 times brighter than the most commonly used GFPs and YFPs. Its excitation maxima is at 506 nm and... things about these proteins is that they are monomeric fluorescent proteins. Fluorescent proteins that...Ai, Hui-wang, et al. "Directed evolution of a monomeric, bright and photostable version of Clavularia ...PMC1698604. Ai, Hui-wang, et al. "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein...

...miRFPs are monomeric and bright Recent series of enhanced monomeric NIR FPs named miRFPs (monomeric infrared... 127 Monomer 394 4.5 37 miRFP709 https://www.addgene.org/79989/ 683 709 78,400 5.4 69 Monomer 192 ...112 Monomer 54 4.5 15  15 miRFP https://www.addgene.org/108409/ 674 703 92,400 9.7 145 Monomer ND ...127 Monomer 700 4.5 90 miRFP713 https://www.addgene.org/136559/ 690 713 99,000 7.0 122 Monomer 980...95,000 10.8 166 Monomer 545 3.7 62 8 miRFP670nano3 645 670 129,000 18.5 387 Monomer 675 4.2 250 9 ...704 93,000 9.9 149 Monomer 1265 4.1 81 miRFP718nano 690 718 79,000 5.6 72 Monomer 885 3.8 33 State-of-the-art... state-of-the-art NIR FPs can be used similar to GFP-like probes by supplying a single gene to cells. ...

... the supplied donor plasmid will express GFP. If there’s GFP expression, presumably the endogenous target...RFP to GFP expressing cells provides a real-time read out of Cas9 activity.  By using the RFP:GFP ratio...in the genome) and an out-of-frame H2B (histone)-GFP reporter into the genome of mammalian cells.  ...puromycin resistance gene linked to an in-frame H2B-GFP. This visual read out allows for easy screening of... the gene NFE2L2, the Madison lab found that H2B-GFP expressing cells exhibited a 2-4 fold increase in... can easily and seamlessly remove the integrated GFP reporter leaving you with just your desired genomic...SRIRACCHA plasmids at Addgene GEMCherry2, an improved monomeric fluorescent protein for assessing CRISPR/Cas9 ...

...et al. developed monomeric FP timers with less propensity to aggregate. These monomeric FP timers (mK-GO... a given timer in many experimental settings. Monomeric fluorescent protein timers Since the production...et al.) were derived from previously developed monomeric fluorescent proteins mKO and mCherry, respectively...events occurring at varied time scales.  Table 1: Monomeric FP Timers and Associated Plasmids from Subach ...neuroendocrine cells revealed by newly developed monomeric fluorescent timer protein." Molecular biology ... PMID: 11408473. 4. Subach, Fedor V., et al. "Monomeric fluorescent timers that change color from blue...Fluorescent Protein Aggregation Catch Up On Your GFP Basics Get Advice on Choosing Your Fluorescent Protein...

...Peptide-assisted genome editing for primary cells Monomeric StayGold variants AAV.CAP-Mac for systemic gene..., Myc and SV40 Nuclear Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral...PMID: 37095348. Breaking up is hard to do: Monomeric StayGold variants By Mike Lacy In 2022, Atsushi...applications. Several labs have been working to produce monomeric versions that break up the dimerization without...Piatkevich and colleagues generated yet another monomeric StayGold variant (Piatkevich et al., 2023). Check...2941917/v1.  Ivorra-Molla, E., et al. (2023). A monomeric StayGold fluorescent protein. Nature Biotechnology...Piatkevich, K., et al. (2023). Bright and stable monomeric fluorescent proteins derived from StayGold. ResearchSquare...

...S65T) in the original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness...generate the strictly monomeric EGFP variant, mEGFP (Zacharias et al., 2002). GFP needs oxygen Chromophore... There are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made...examined on a case-by-case basis. GFP is sensitive to acid The chromophore of GFP can exist in a protonated and...its acid tolerance is maintained in cells.   GFP is big GFP is a 28 kDa protein that resembles a cylinder...2013; Hilaire et al., 2017). GFP can only be used for tagging proteins GFP fusion proteins are made by ... a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP fluorescence overlaps...

...the limitations is the size of GFP: ~240 amino acids or about 28 kDa. GFP and its homologues have a beta-barrel...potential issues, GFP has been used successfully in countless fusion proteins. Choose monomeric proteins and...and shape matters The green fluorescent protein (GFP) from Aequorea victoria and its variants are genetically...interference is known as steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding sites... an inert light emitting module, any tendency of GFP to homodimerize should be avoided because it will...Keep the linker sequence short In the early days of GFP applications, many were concerned with steric hindrance...fluorescent proteins derived from Aequorea victoria GFP sticks out and can be considered a linker (figure...

...a gene of interest fused to the N-terminus of monomeric Cerulean mCerulean C1 Cyan Mammalian Express...a gene of interest fused to the C-terminus of monomeric Cerulean mTurquoise2 Cyan Mammalian Constructs...a gene of interest fused to the N-terminus of monomeric Venus mVenus C1 Yellow Mammalian Express a gene... gene of interest fused to the C-terminus of monomeric Venus pCEP4YPet-MAMM Yellow Mammalian Expresses...SYFP2 Clover Green Mammalian Expresses Clover (a GFP variant) commonly used with mRuby2 pLSSmOrange-...