... neurons and show that smFPs fusions provided better labels than fusions to 3 or more HA tags. 4. High-resolutions...synapses for instance. Also these tags need to be fused to scaffold proteins in order to be expressed stably...contrast to epitope tags, FPs are bigger and their fusion to a POI can affect protein folding and protein-protein...proteins (smFPs). smFPs are literally composed of FPs fused to multiple epitope tags. Indeed, to create the ...difficult to distinguish using standard antibodies. By fusing N-cadherin to smFPs the Looger team was able to...

...complexes, scientists can simply express a bait protein fused to BirA* in cells and then incubate those cells ...and allows scientists to more selectively target fusion proteins to specific locations in the cell.   Find...increased with higher expression levels but for many fusion proteins this would lead to protein/organelle aggregation... is an in vivo technique where two proteins are fused to complementary reporter proteins that can only...molecule that can reassemble to restore activity when fused to two interacting proteins (Schopp et al., 2016...Roux, Kyle J., et al. "A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins...

...epitope tag fusions to a protein of interest. The plasmids used to generate these fusions all use the ...of functions via different fusions. Some applications of PUF protein fusions reported in the literature...leader sequence. Using a variety of GFP and RFP fusions as their test genes, the researchers at the BIOFAB...little as 3 days. Abil et al., 2014 show that you can fuse custom PUF proteins to a translational repressor...

...45 kDa tag which can be purified (along with its fusion protein) with amylose resin. The MBP tag, although...Unlike MBP, the GST tag can be cleaved off from the fusion protein while it is still bound to the glutathione...proper folding of the tag, which prevents insoluble fusion proteins from being purified by GST (maybe try ...ions (Nickel, Cobalt, Copper, etc.), allowing His fusion proteins to be purified by single step metal affinity...affinity chromatography. Purification of His fusion proteins from E. coli is generally more straightforward...

...stable scaffold, part epitope binding region, all fused to a fluorescent protein. “Frankenbody,” like the...variable fragments (scFvs) (Inbar et al., 1972) are fused to mature fluorophores, and can bind and light up...scaffolds specific to different targets, the scientists fused them to sequences called complementarity determining...of interest tagged with the HA epitope, the lab fused a fluorescent protein to the hybrid anti-HA scFvs...sensitive cells such as living neurons, and can be fused to other fluorescent proteins and tags including...

...protein that they are fused to. The researchers constructed 22 different mTFP1 fusions to both the C- and...readout, respectively. mNeonGreen can be used as a fusion tag for a wide range of applications from traditional...mCitrine and mOrange or for live cell imaging of fusions and FRET reporters. mTFP1 is also insensitive to...termini of the fluorescent protein and found that the fusion protein localized to their expected locations. ...

...of plasmids each containing an anti-GFP nanobody fused with a different functional element (tyrosine sulfation...described here can be used to target any protein fused with GFP, YFP, mCerulean, or other related fluorophores... He made use of catalytically inactive dCas9 by fusing it to an engineered peroxidase, (APEX) that labels...plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed from a Tet/Shield1 inducible...the 20 nm radius surrounding the dSpyCas9-APEX2 fusion.  This Sontheimer lab used this technology, which...

...easier - now, one can generate a fluorescent protein fusion, transcriptional reporter, and loss-of-function...template plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The repair template and Cas9-sgRNA plasmid...loss-of-function allele as well as a promoter-FP fusion for the gene of interest. L1/L2 larvae from these...phenotype. These animals contain a fluorescent protein fusion for the targeted gene. After initial testing, Dickinson...genome. Although they tested fluorescent protein fusions, this system could be used to make many other genome...

...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...

...primer editor protein consists of a Cas9 nickase fused to a reverse transcriptase. The Cas9 nickase part...examine the parts in more detail. The prime editor: A fusion between Cas9 and reverse transcriptase To decrease...editing would introduce into the cell, the team fused the M-MLV reverse transcriptase (RT) with the Cas9...editor (PE). They found that orientation matters: fusing the RT to the C-terminus of the Cas9 nickase resulted...thermostability. What came out on top? The Cas9 nickase fused to a pentamutant of M-MLV RT. They called this system...

... 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...

...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...