...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...

... 204223 CMV-Halo-10-PFN1-Y6D PFN1 Halo, Tev CMV ALS Jessica Henty-Ridilla 204224 CMV-Halo-10-PFN1-R88E...R88E PFN1 Halo, Tev CMV ALS Jessica Henty-Ridilla 204225 CMV-Halo-10-PFN1-G118V PFN1 Halo, Tev CMV ALS Jessica... 161767 FUS_Halo_N_allele_400 FUS Halo ALS Michael Ward 161768 FUS_Halo_C_allele_400 FUS Halo ALS Michael...Melcher 178115 ANG_Halo_C_allele ANG Halo ALS Michael Ward 178116 PRNP_Halo_C_allele PRNP Halo Dementia Michael...178117 PFN1_Halo_C_allele PFN1 Halo ALS Michael Ward 178118 CHCHD10_Halo_N_allele CHCHD10 Halo ALS Michael...178119 UBQLN2_Halo_C_allele UBQLN2 Halo ALS Michael Ward 178120 SIGMAR1_Halo_C_allele SIGMAR1 Halo ALS Michael...178121 APOE_Halo_C_allele APOE Halo Alzheimer's Michael Ward 178122 TREM2_Halo_N_allele TREM2 Halo Alzheimer's...

...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...

...enhancements such as: Human codon optimization Example: Halo Increased photocurrent amplitude Examples: eNpHR...strain Shark), used to silence neuronal activity 632 Halo/NpHR Light-driven inward chloride pump from Natronomonas...function. TULIPs Tunable light-inducible dimerization tags (TULIPs), plasmids allow cloning of protein coding...sequences with GFP-LOVpep, cpPDZ, ePDZb and ePDZb1 as tags. pDawn/pDusk Blue-light photoreceptor for light-...plasmids . Halorhodopsins Halorhodopsins are light-gated inward chloride pumps isolated from halobacteria. Wild-type...Jaws Browse Halorhodopsin plasmids . Archaerhodopsins Archaerhodopsin-3 (Arch) from Halorubrum sodomense...variant 517 Halorhodopsins Jaws Red-shifted, light-driven inward chloride pump from Haloarcula (Halobacterium...

...NanoLuc® with either SNAP‐tag or HaloTag7 tag in which labeling of the self‐labeling tag with a fluorophore ...biosensor that utilizes Renilla luciferase and a Halo tag to assay mRNA decay in real time. pKC-4.04, pKC...and Renilla luciferase are less-than-ideal protein tags due to their large size. NanoLuc® Luciferase is ...Lentiviral expression of firefly luciferase with a V5 tag Kevin Janes 83281 pAAV-CAG-FLuc Firefly CAG AAV expression...Mammalian expression of Nanoluc with a N-terminal Myc tag Erich Wanker 124701 pLenti-PGK-Venus-Akaluc (neo)...Lentiviral expression of Nanoluc with a N-terminal Myc tag Erich Wanker 115352 pFL-SV40 Firefly SV40 Mammalian...systems. Each type of luciferase has advantages and disadvantages depending on the application. Firefly...

...analogs Mammalian TAG Huiwang Ai 160041 pRaGE Pyl TAG GFP Y35TAG PylRS M. mazei Bacterial TAG Lital Alfonta...pPaGE Pyl TAG FliC T248TAG PylRS M. mazei Bacterial TAG Lital Alfonta 160377 pDule-Mb haloTyrRS C6 C6 HaloTyrosine...nitroY/haloY-F5 PylRS M. alvus 3-nitrotyrosine (3-nitro-Y), 3-halotyrosine (3-halo-Y) Bacterial TAG Ryan...DiZHSeC Mammalian TAG P. Chen 92047 pCOTS-pyl-GFP(35TAG) PylRS M. mazei Cyanobacterial TAG Lital Alfonta ...Mammalian TAG Kensaku Sakamoto 197099 pIYN3 TyrRS M. jannaschii halogenated tyrosines Bacterial TAG Kensaku... Bacterial TAG Peter Schultz 48696 pANAP AnapRS E. coli fluorescent AA, Anap Mammalian TAG Peter Schultz...coli pBof Mammalian TAG Huiwang Ai 68292 SepOTSλ SepRS9 E. coli Sep Bacterial TAG Jesse Rinehart 71403...

...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...

...with endogenously-tagged cellular structures. Addgene ID Plasmid Allen Institute ID Tag Protein Structure...has deposited plasmids to generate fluorescently tagged proteins of key cellular structures in human cell...You May Also Like... CRISPR Guide CRISPR Protein Tagging Fluorescent Protein Guide Subcellular Localization... publicly available collection of fluorescently tagged, human, induced pluripotent stem cell (hiPSC) lines...researchers will be able to generate fluorescently tagged proteins of key cellular structures in human cell...plasmids, check out Plasmids for Endogenous Gene Tagging in Human Cells on the Addgene Blog. Allen Institute...TTN-mEGFP AICSDP-45 mEGFP Titin Sarcomeres (M-line tag) 114413 MYL7-mEGFP AICSDP-46 mEGFP MLC-2a Sarcomeric...

...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...