...sample. The second sensor is a fusion protein consisting of sfGFP fused to a His-tagged ABC transporter...exposed to ultraviolet light. To do so, she generated fusion proteins in which Dronpa domains flanked either...either side of an enzyme’s active site. In these fusions, ultraviolet light exposure caused oligomerization...iterations of FLiPs only used N- and C-terminal dronpa fusions. With these various designs, Xin has been able ...

...Moshe Szyf at McGill University. Some enzymatic fusions are godsends. Tethering dCas9 to the histone acetyltransferase...where they were absent before. But other enzymatic fusions, such as dCas9-TET, can, like the iron ball-and-chain...notoriety – as a tool of choice in the field: dCas9 fused to the enzymatic domain of TET1 or TET2 (Morita ...by dCas9 alone, without TET (or any demethylase) fusion. This may seem counterintuitive: how can a DNA-...–peptide repeat and scFv–TET1 catalytic domain fusions. Nat Biotechnol 34, 1060–1065. https://doi.org/...

... own. When the two fragments come together, they fuse and fluoresce. The team engineered C. elegans cell...2020.07.02.185249 Predicting cell cycle length with a histone-fused fluorescent timer By Kate Harten DeMaio Studies ...Shangqin Guo and her colleagues at Yale University fused a color-changing fluorescent timer (FT) protein ...hours in primary mouse embryonic fibroblasts. This fusion protein can be used to determine cell cycle length...lentiviral preps at Addgene hSyn1-SIO-stGtACR1-FusionRed is a Cre-dependent, soma-targeting inhibitory ...

...purified by centrifugation. Gag-Cas9 fusion protein The MLV Gag protein fused to a Flag-tagged SpCas9 is incorporated...transcriptional activator dCas9-VPR, if these variants are fused to the MLV Gag in place of SpCas9. gRNA A gRNA targeting...association with the the Cas9 portion of the Gag-Cas9 fusion protein. A single Nanoblade particle can be loaded...space surrounding the zygote, where they can then fuse with the target cell membrane, and transfer their...

... that they only facilitate degradation of their fused protein under certain conditions (typically the ...novel degrader of the 12 kDa protein FKBP12F36V. By fusing FKPB12F36V to your protein of interest, it can ...molecule which induces dimerization of the FKPB12F36V fusion protein with the endogenous CRBN E3 ligase complex...complex which will polyubiquitinate the fusion protein. This will target the protein of interest to the ... molecules induce dimerization of an FKBP12F36V fused target protein-of-interest and the CRBN E3 ligase...

... perfectly capable of synthesizing fluorophore fusions on their own. Subsequent advances in fluorescent...moving from transient overexpression of protein fusions to gene-edited cells can decrease the number of... (1) a genetically encoded “tag” expressed as a fusion with your favorite protein and (2) a synthetic ...small-molecule labeling approaches can provide a fresh infusion of photons for your imaging experiment. Innovative...Resources on Addgene.org Generate Fluorescent Protein Fusions Request Plasmids from the Michael Davidson Collection...

...constantly being developed! Fusion tagging: One of the most common uses, GFP can be fused to the N- or C-terminus...of empty backbones for constructing fluorescent fusions. Transcription Reporter: Placing GFP under the ...overlapping excitation/emission spectra are used; one fused to each protein or domain being tested. Find FRET...interactions. In this case, two portions of EGFP are fused to the proteins of interest, and when they come ...

... of E. coli tRNA adenosine deaminases (ecTadA), fused it with dCas9, and transformed it into E. coli harboring...and tomato using a CRISPR-Cas9 cytidine deaminase fusion." Nature biotechnology 35.5 (2017): 441. PubMed... wheat and maize with a Cas9-cytidine deaminase fusion." Nature biotechnology 35.5 (2017): 438. PubMed...Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A." Nature biotechnology...rice and wheat using a Cas9-adenosine deaminase fusion." Genome biology 19.1 (2018): 59. PubMed PMID: ...

...catalytically dead Cas9 (dCas9) alone or a dCas9 fused to the omega subunit of RNA polymerase, the lab   ...for CRISPRi and CRISPRa using RNAP-Omega-dCas9 fusions. Here’s some others. SoxS based CRISPRa In the ...The authors were able to engineer a dCas9 variant fused to bacterial enhancer binding protein required for...editing. This process is generally accomplished by fusing an enzyme capable of converting one base to another...identified a cytidine deaminase, PmCDA1, that could be fused to dCas9 and efficiently introduce cytosine-to-thymine...glycosylase inhibitor and degradation tag on the fusion protein to limit the activity of the system as ...

...binding scFv that’s fused to GFP. When the multiple copies of the scFv-GFP fusion bind the SunTag scaffold...smaller than antibodies. scFvs are often genetically fused to other proteins, which has led to their application...detection probe. It contains an HA-binding scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody...brings T cells to a tumor site. ScFvs can also be fused to cellular toxins, radioisotopes, cytokines, and...

...and deposited versions with both PDGFR and NGR fusions for membrane display, they recommend the NGR vectors... Figure 4: STITCHR consists of a nicking Cas9 fused to a shortened R2Tocc transposon protein, and can...the transposon RNA homology both help direct the fusion construct to the new genomic locus. Image courtesy...After minimizing the system's size, the scientists fused the retrotransposon protein R2Tocc to a Cas9 nickase...better kinetics and less background labeling. By fusing miniTurboID with dCas9 to create TurboCas, the ...

... expression. However, one caveat of FP-protein fusions (FP-chimeras) is that they undergo normal protein...their continuous excitation. By generating PA-FPs fused to intracellular proteins, scientists can track ...as organelle trafficking and dynamics (fission, fusion, etc). For example, by engineering PA-GFP tagged...Youle Lab was able to visualize and quantify the fusion dynamics of mitochondria in healthy and apoptotic... individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis...

...generating GFP fusion proteins. From considering protein size and shape, linker sequences, and fusion location...they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection by Joachim...control and flexibility to CRISPR editing. Using a fusion between Cas9 and reverse transcriptase and a prime...

...desired (Figure 1). The probe perfuses liquid, which causes the passive diffusion of DA across the semipermeable...extracellular fluid into the perfused liquid. The probe then collects the perfusion liquid for quantitation...

... of a quickly-degraded destabilized GFP variant fused to a 4X CDRE repeat. In addition to showing that...2Fmic2020.04.713 Truncated TALE-fluorescent protein fusions as a DNA staining dye Article contributed by Jennifer...Kyubong Jo's lab generated fluorescent protein fusions with tTALE to “stain” DNA. TALE proteins can bind...structural deformation or photo-induced damage. These fusions can also function in high salt conditions - up ...

...that bind to cobalamin-fused fluorescent dyes; binding of the cobalamin fusion to the riboswitch results...containing the growth promoting domain. Each half was fused with one of the LOVTRAP domains. When both halves...quantification of base editors that utilize dCas9 fused to cytidine deaminases like APOBEC. To achieve this...

...protein. The prime editor consists of a Cas9 nickase fused to a reverse transcriptase. The Cas9 nickase is ...strand. The first iteration of prime editor simply fused the wild-type Moloney Murine Leukemia Virus (M-MLV...thermostability. What came out on top? The Cas9 nickase fused to a pentamutant of M-MLV RT. They called this system...3′ tail of pegRNAs (Yan et al., 2024). The team fused La to the C-terminal end of PEmax to produce PE7...pegRNA stability ~ ~ ~ ~ ~ ~ ✖ Fuse RNA-binding protein La to the prime editor Improve...

...demonstrate that they can successfully create GFP fusion proteins with a variety of genes across the genome...dehydrogenation). MitoTimers are composed of a Timer protein fused to the mitochondrial targeting sequence of cytochrome...perform multiple site mutagenesis and create complex fusion or recombination vectors. The efficient and flexible... is a monomeric, 27 kD genetic tag which can be fused to proteins of interest for EM imaging, or targeted...colleagues, FokI-dCas9 expresses Fok1 nuclease domain fused to catalytically inactive Cas9 DNA-binding domain...single reaction, while retaining only a defined 4bp fusion site in between. The MoClo system is comprised...

...domain.  How do these components interact? When fused together to form an OptoNB, the AsLOV2 domain acts...Since Ras is a membrane localized GTPase, the team fused a mCherry OptoNB to the catalytic domain of the ...that activates Ras. They expressed this OptoNB-SOS fusion protein in cells that expressed membrane-localized...activity Read about RANbodies, reporter nanobody fusions Learn more about using the secondary nanobody toolbox...

...its target.  They name their most potent Acr-LOV fusion protein inhibitor CASANOVA for “CRISPR-Cas9 activity...works with the S. pyogenes Cas9, dCas9-effector fusions, and xCas9, and requires no additional chemicals...plasmid tools which includes plasmids encoding a fusion between cPYP and tgRFP (PLL7.0 tgRFP cPYP), and...or dark- state of the photoswitchable proteins) fused to mVenus linked to a mitochondrial localization...

... delivery into mammalian cells, especially when fused to effector proteins for gene activation or gene...the new system was able to activate expression by fusing dCas12f to VP64. Read the paper in Nature Biotechnology...combines an endogenous Gag homolog, cargo RNA, and fusogen in a modular system for delivering gene editing...active Cas9 enzyme. PE-Cas9 comprises an active Cas9 fused to the reverse transcriptase (RT), which induces...

... Early efforts tested whether or not FP-tubulin fusion proteins could properly form microtubule networks...networks in cells. Other efforts tested whether FP fusions to connexin localized correctly to gap junctions...OSER assay with a number of fluorescent proteins. Fusions of CytERM with mTurquoise2 and dTomato, which are... localization and biological properties of new fusions can be compared with mEGFP or mTurquoise2, which...

... cytoplasmic contents. Autophagosomes that have fused with the vacuole or a lysosome are trickier to identify... are degraded or released once an autophagosome fuses with the vacuole or lysosome. Rosella fluorescent...They are connected by a 9 amino acid linker that’s fused to the C-terminus of DsRed.T3. See Table 1 for a...Sargsyan et al. It is also possible to make Rosella fusions and thereby monitor autophagy of your protein of...

... the second type of these GEVIs: a FP-fused rhodopsin. FP-fusion to rhodopsin resulted in electrochromic... Simply put, VSD-based sensors consist of a VSD fused to a single fluorescent protein (FP). This class...imaging.   Figure 1: Ace-mScarlet is a fusion between the voltage-sensitive inhibitory rhodopsin...

...exact entry for your favorite gene fused to an epitope tag or fusion protein. If your sequencing primer...An alignment omitting repeated regions can be confusing, such as when you attempt to verify the starting...primer was chosen to confirm a tag or fusion protein is in-frame, we recommend using blastx with the "Align...

...ectopically expressed in most organisms. Furthermore, fusing FPs to another protein usually does not affect ...and 2) to visualize the localization of the FP (fused to the protein of interest), thus tracking the localization...used to determine the proximity of two proteins fused to the halves of the split FP. This technique is...This may affect the biological function of the FP-fusion protein. Therefore, it is recommended to use monomeric...

...has focused on transcriptional activation using a fusion to a transcriptional activator such as VP64 (Gilbert...transcriptional activators (Qi et al., 2013). However, the fusion of dCas9 with a protein tag allows for the isolation... DNA fragment (Fujita et al., 2016). 3xFLAG-tag fused to dCas9, along with a biotinylated gRNA, were successfully... (Liu et al., 2017). This system utilizes dCas9 fused to a biotinylation tag expressed on a plasmid, allowing...

...modular approach combines an interchangeable set of fusogenic and targeting components to reprogram delivery...Modular features of DIRECTED particles where the fusion and targeting components are interchangeable. Image...targetable cell types. DIRECTED is compatible with fusogens from different viral families and can be used ...transcriptional history of a cell. Lin et al. generated a fusion protein called iPAK4 that assembles into a slow-growing...

...MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study) are captured on...programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase — that can drive lasting...silencing, which can be toxic, CHARM uses a cofactor fused to a histone tail to recruit and stimulate the endogenous...multiplexed targets.   Figure 4: CHARM uses a fusion of the DNMT3L cofactor (D3L) with a histone tail...

...expression, measured using fluorescent protein fusions In addition to visible light, flow cytometers can...Which Fluorescent Protein Should I Use? However, fusion proteins tag your protein of interest with a fluorescent...- or C-terminus of the protein. One downside of fusion proteins is that the tag can alter the structure...get around the challenges of fluorescent protein fusions by binding to the protein in its native state.  ...

...experiments. However, remember that, if you make an FP fusion, you are inserting about 20-25 kDa of protein mass...to this bulky insertion are inevitable. Most FP fusions for FRET studies are at the N- or C-terminus of... protein of interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition, since these...FRET Browse Empty Plasmids to Fluorescent Protein Fusions Check out Our Fluorescent Protein Guide ...

... any tags or fusions that are on your insert. Typically you would list any tag or fusion protein in the... pBACKBONE-Flag-hGene If there was also an EGFP fused to the C-terminal of your insert you would list ...

...After testing dCas9 fusions with transcriptional repressors, they found that fusing dCas9 to an Mxi1 repressor...covalently attached to the Cas9-gRNA RNP complex via the fused HUH endonuclease PCV. This increases HDR efficiency...

..., Georgieva et al. examined the effects of GFP fusions to inntags or traditional FLAG, HA, and Myc tags...untagged GFP overexpression, while the rest of the fusions had some negative effect on these parameters. IT5... cell morphology was observed with IT5 and IT6 fusions. In contrast, Flag and HA tags had especially detrimental...

...from the Zhang Lab. It's name, though a little confusing to the uninitiated, contains much of the information...directed by a gRNA. 2A-Puro In this plasmid, SpCas9 is fused to the puromycin resistance gene through a 2A peptide...the 2A peptide on its C-terminus, SpCas9 is also fused to a 3X FLAG tag at its N-terminus. This tag can...

...they identified included gene amplifications and fusions, missense mutations, and indels (Ng et al., 2018...carrying a single gene with a defined mutation or gene fusion. Plasmids carrying the corresponding wild-type ...Mutations in Cancer.” Of all the mutations and fusions represented in this collection, approximately 80%...

...genome editing by fusing Cas9 to a mutated human estrogen receptor. This protein fusion stays in the cytoplasm...vector to create knock-ins of fluorescent proteins, fusion proteins, or transcriptional regulators. In this...

...by using the DNA cutting activity of a Protein A fused micrococcal nuclease (MNase) to specifically isolate... against the protein of interest. The Protein A fused MNase is then added and Protein A binds Immunoglobulin...protein interactions Learn about other types of fusion proteins Study chromatin interactions with CAPTURE...

... for Goldengate cloning   Design primers for In-Fusion Cloning Design primers for Gibson Assembly and ...restriction enzyme cloning, Gibson assembly, or In-Fusion cloning.  NEB Ligation Calculator: This tool works... from five different insert:backbone ratios. In-Fusion cloning molar ratio calculators Lab math calculators...

...an exon of a gene to generate a transcriptional fusion. This may disrupt the endogenous function of the...the gene product resulting in a transcriptional fusion, much like promoter traps. While this system does...disrupt the gene it monitors due to transcriptional fusion.   Examples of promoter, enhancer, and gene...

...biosensors contain a single fluorescent protein fused to a domain that senses the concentration of a given...an example. Many calcium biosensors include GFP fused to calmodulin (CaM) and the chicken myosin kinase... an intracellular or extracellular molecule? By fusing a biosensor to an organelle-specific targeting ...

...available – you can always add an epitope tag or a fusion protein. Mammalian vectors One of our most popular... cells. The addition of an in-frame fluorescent fusion protein will allow you to visualize YGOI in vivo... offer a collection of vectors carrying several fusion proteins. Pick your promoter, selection cassette...

...binding protein known as GltI. When the resulting fusion protein binds to glutamate, conformational changes...original iGluSnFR in the pMinDisplay backbone is fused to a specific peptide segment (PDGFR TMD) that anchors...Aggarwal et al… Figure 4. PDGFR is a C-terminal fusion to the PDGFR transmembrane domain in the mammalian...

...conditionally and reversibly degrade proteins of interest fused to a degron, a domain that induces degradation. ...In addition to the engineered SpCas9, they also fused the SpCas9-NG with the activation-induced cytidine...light-activated-dynamic-looping (LADL) system. The group fused dCas9 to a truncated CIB1 protein from Arabidopsis...

...sites to tether many copies of the FP-coat protein fusions to their reporter mRNAs. Using translation inhibitors...to the plasma membrane; this sequence prevents diffusion of the mCherry labeled mRNA and keeps it in a ...SunTag, a synthetic scaffold that can recruit GFP fused to the SunTag-specific antibody scFv. Each time ...

...diversification by targeting a site of interest with dCas9 fused to the Activation Induced Cytidine Deaminase (AID...two key components: the error prone PolI3M that’s fused to a nicking Cas9 (nCas9). Similar to mutagenesis...consists of an error-prone polymerase (PolI) that is fused to a nicking Cas9. A guide directs Cas9 to a site...

...producing monoclonal antibodies is from a hybridoma, a fusion between an antibody-producing B cell and a myeloma...cells from an immunized animal, those B cells are fused to myeloma cells at a 1:1 ratio and then cultured...medium called HAT. This medium selects against unfused myeloma cells and antibody-producing B cells that...

...beta-arrestin-TEV protease fusion to a receptor-transcription factor fusion. TEV protease cleave the transcription... et al., 2017). In this technique, the receptor fused to APEX2 catalyzes the biotinylation of proteins...

... mEGFP, mRuby2, iRFP713, SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies include mEGFP,...Leeson The large size of CRISPR-Cas effectors and fusion proteins, such as Cas9 and Cas12a, creates challenges...from Xiaoshu, et. al. 2021.   Created by the fusion of Cas12f with a transcriptional activator, CasMINI...

...proteins into one particle within the cell, the lab fused the red fluorescent protein (RFP) to a self-assembling...microcompartment protein. The expression of this fusion is under control of the arabinose-inducible promoter... with arabinose to begin expressing the SAP-RFP fusion. Then, the cells are washed to stop the production...