...degron tags. What is a degron tag and why should I use one?   Degron tags are a class of protein tag which...amenable to being tagged, e.g. tagging some proteins can affect their function, the SMASh tag self-cleaves ...). Choosing a degron tag Are all tags created equal? No! Choosing the ‘right’ tag does depend a lot on...Protein Tags Plasmids for Endogenous Gene Tagging in Human Cells Antibodies 101: Epitope Tags  ...) a SMASh-tagged protein and 2) asunaprevir (small molecule). dTAG The degradation tag (dTAG) system functions...degradation. Like several other tags described here, the dTAG uses a small molecule, a tag, and the endogenous cellular...machinery to function. The dTAG is the smallest tag reviewed here, in regard to total tag size, which can also...

... with containing chemical tags such as the SNAP-tag, Halo-tag, and CLIP-tag (Kohl et al., 2014) (Addgene... et al. "Ultrafast tissue staining with chemical tags." Proceedings of the National Academy of Sciences111.36... circuits with fluorescent proteins and chemical tags." eLife 7 (2018): e40350. PubMed PMID: 30454553....

...T2A-TurboGFP-PEST), and small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The Foerstemann lab developed...types of fluorescent (EGFP, tagRFP, mCherry) and nonfluorescent (Halo, SNAP) tags, a selectable marker (floxed... scarless tag insertion.   In addition to the obvious advantages SapTrap offers for tagging a single genetic...between the tag and homology arms). Digestion of the donor plasmids with SapI allows the tag, selectable... inserting a tag at multiple target sites. SapTrap vectors can be easily modified to tag hundreds of genes...insertion tagging), developed by the Hornung Lab, allows users to easily create C-terminal tag fusions ...endogenous protein tagging, please visit our website CRISPR/Cas Plasmids - Protein Tagging. Beyond simply...

...Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag. Biochemistry, 60(33), 2560–2575...genetic tag on the protein of interest. Table 1: Summary of popular self-labeling tag systems Tag Size...than short peptide tags like tetracysteine (Jing & Cornish, 2011). HaloTag and SNAP-tag are the most popular...application, SNAP-tag, CLIP-tag (both 182 aa), and eDHFR (159 aa) are smaller than HaloTag (295 aa) – for...flexibility of self-labeling protein tags. The basics These tags most often consist of an enzyme that ...resorufin). SNAP-tag: In 2003, Kai Johnsson’s lab developed a self-labeling fusion tag based on the human...later became known as SNAP-tag. There is also a fast-reacting version of SNAP-tag called SNAPf. eDHFR: The...

... of N- and C-terminal tags (including His, S-tag, OneStrep, CBP, Trx, GST, Halo, MBP, NusA and SUMO) for...electron microscopy (EM) tag providing superior subcellular localization of tagged proteins compared to traditional...promoter, N-terminal tag, coding sequence of the gene of interest, C-terminal tag, plant terminator, and...of an isopeptide bond between 2 peptide tags, SpyTag and KTag. The group demonstrated the use of the SpyLigase...expression levels. APEX2 is a monomeric, 27 kD genetic tag which can be fused to proteins of interest for EM...same animal. Flipbow additionally incorporates SUMO tags in the FP sequence for separation from Cre-based...levels to be directly compared among the different tags To clone a sequence or gene of interest into the...

...would be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme variant tag that covalently...several advantages to using these self-labeling proteins over traditional fluorescent protein tags, such...endogenous target, and thus no genetically encoded tag is required. These dyes can also be attached to an...ligands have more flexibility due to the affinity tag they typically bind to. Stability of fluorescence...experimental labeling. Size is also a factor. Bulkier tags, such as GFP, may interfere with protein expression...fluorescent experiments, such as using fluorescently tagged ligands or intermolecular FRET for ligand-receptor...should possess high specificity to ensure accurate tagging. In Fluorescence Resonance Energy Transfer (FRET...

... small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the tag can recruit multiple...SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies include mEGFP, mCherry, and HaloTag. ...b) Schematic of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of...blotting, and more. Given the popularity of FLAG and HA tags, you may not even need to clone new constructs to.... The proteins, which are purified via histidine tags, are optimized for SDS-PAGE gels and can be detected...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure...

...experiment.  Highlighted Split-Fluorescent Protein Tags Browse the table below, and click the Article link...cellular proteins with split fluorescent protein tags. Tamura R, Jiang F, Xie J, Kamiyama D. Commun Biol...most-requested GFP(1-10) and GFP(11) Versatile protein tagging in cells with split fluorescent protein. Kamiyama... monomeric near-infrared fluorescent proteins as tags and biosensors for multiscale imaging. Shcherbakova... Fluorescence-activating and absorption shifting tag (FAST) for use with green-yellow or orange-red fluorogen...

...other fluorescent proteins and tags including mCherry, HaloTag and SNAP-tag for multi-color imaging. Find...observed colocalization of GFP tagged HA frankenbody with an mCherry, 4xHA tagged histone protein (H2B) in ...could also bind a 1xHA tagged histone protein as well as a mitochondrial protein tagged with spaghetti monster... the 10xHA tag increased the fluorescent signal to noise ratio five fold over the 1xHA tag.  Lastly, the...use GFP to image nascent proteins. In addition GFP tags are large, dim, and permanently attached to a protein...limitations, scientists have co-opted antibodies for tagging purposes. Different antibodies bind different protein... called epitopes and scientists can use these to tag their proteins of interest. Antibody based probes...

...concept include: Self-labeling tags (e.g., SNAP-tag, HaloTag, TMP-tag) – These widely used systems consist...strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion with your favorite protein...fluorophore-contianing “ligand” that binds to the tag. Like most good ideas in biological imaging, the ...genetically encoded tetracysteine (Cys4) peptide tag could be used to label proteins in cells (Figure ...consist of a genetically encoded enzyme variant tag that reacts specifically and irreversibly with a small... attachment of a fluorophore ligand to a peptide tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine...paclitaxel. Unlike other systems, no genetically encoded tag is required as the small-molecule binding motif targets...

...lentiviral vectors to stably or inducibly express tagged iPAK4 from different promoters, specific time points...plasmids here! Lin D., Li X., et al. (2023). Time-tagged ticker tapes for intracellular recordings. Nature...externally-timed bands of color with HaloTag-iPAK4 by applying different HaloTag dyes and showed they could record.... B) HEK cell with an iPAK4 fiber labeled with HaloTag-iPAK4. Scale bar 10 µm. C) Images of iPAK4 fibers...fibers in cultured rat hippocampal neurons. HaloTag dyes are switched from yellow to red at 0 h and back...

...fact, NLuc is bright enough to detect endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally..., NLuc is very inviting for endogenous protein tagging as it is only 19 kDa. An example is the CRISPaint-NLuc...2016) CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nature Comm...membrane-inserted voltage sensing domain (LOTUS V; Luminescent Optical Tool for Universal Sensing of Voltage). With...information. Many of our technology platforms like HaloTag® Fusion Protein, codon-optimized Firefly luciferase...available through Addgene (Plasmid #83926). Live-Cell Voltage Sensor  Ingaki and colleagues (8) were interested... al. (2017) Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of...

...allow for noninvasive live imaging with fluorescent tags and activation of light sensing proteins. Furthermore...become a popular model organism because their larval stage lends itself well to studies of neuroscience. The..."Optical control of zebrafish behavior with halorhodopsin." Proceedings of the National Academy of Sciences...

...Addgene is certainly going to have to find ways to tag and curate reagents from these emerging organisms... a different protein. Did you know that most cephalopods die after they lay their 1 clutch of eggs? Propagation...

...Voltron2 voltage sensors By Brian O’Neill New viral vector preps expressing the Voltron2 voltage indicator...-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting microglia The...earlier Voltron indicators (a fusion of Ace2N and HaloTag, Figure 4), they determined that an A122D mutation...ratio and better ability to record subthreshold voltage fluctuations, while having high fidelity during...neurons. The same A-to-D mutation in the related voltage indicator Ace2N-mNeon also led to similar improvements...negative-going sensors (with fluorescent signal at baseline voltage), the investigators found that administering these...changing the dye that becomes conjugated with its HaloTag domain.  Find Voltron2 plasmids here! Abdelfattah...

...expressed from the same cassette is sometimes advantageous, particularly when only a portion of the plasmid... mRNA. However, some bicistronic vectors take advantage of an element called an Internal Ribosome Entry...bicistronic vectors; however, they do have some disadvantages. These elements are quite large (500-600 bp)...vector.3  2A Peptides To overcome some of the disadvantages of the IRES element, scientists have adapted...segment of the 5' nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes...

...guide sgRNA-Sirius-8XMS2 which will recruit the HaloTag with red fluorescent ligands to the loci of interest...

...and subsequent firing of an action potential. Halorhodopsins (HR) are green/yellow-light sensitive chloride...promoter of the cre recombinase while still taking advantage of the robust expression via the promoter in the...stereotaxic frame with a digital readout. The advantage is you can “zero” your coordinates at bregma which...engineered to express various opsin genes [7-9]. The advantage of using these lines is, if one is crossed with...