...combines components of SAM and SunTag activators, fusing the epitope tag of SunTag with the P65-HSF activation...SAM. This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou..., Qi LS, Weissman JS, Vale RD (2014) A Protein-Tagging System for Signal Amplification in Gene Expression... for CRISPR activation, research in the area has continued to develop. These newer methods have not yet... of rearranging chromatin structure. This method has shown higher activation levels than SAM in some loci...popular activation methods (VPR and SunTag) (Chavez et al., 2016). SunTag Description Rather than using a ...activation levels are similar to that of SunTag. An advantage to this method compared to other notable...

...contain a variety of N- and C-terminal tags (including His, S-tag, OneStrep, CBP, Trx, GST, Halo, MBP, ...promoter, N-terminal tag, coding sequence of the gene of interest, C-terminal tag, plant terminator, and...Ting’s lab has enabled progress on both fronts – as a genetic electron microscopy (EM) tag providing superior...of an isopeptide bond between 2 peptide tags, SpyTag and KTag. The group demonstrated the use of the SpyLigase...vector for bacterial mutagenesis or gene targeting Victor de Lorenzo's lab has engineered a modular...Building off their SpyTag/SpyCatcher system for protein-peptide locking, Mark Howarth’s lab has developed a ... superior subcellular localization of tagged proteins compared to traditional fluorescence microscopy,...

...KB Fujii iChIP/enChIP to purify genomic DNA FLAG tagged dCas9 PDF, 110 KB Goldstein Nematode: gRNA design...PDF, 109 KB Mendenhall and Myers Mammalian: FLAG tagging endogenous proteins pFETCh_Donor ; additional HDR...cloning in gRNA: pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) PDF, 49 KB Vosshall and Matthews (Link opens...

... cells. Flexible tagging: Choose untagged or add a versatile N- or C-terminal HA tag. Selectable and stable...EGFP into pAG Lenti CMV N-HA Puro (Addgene #236079) to create pAG Lenti CMV HA-EGFP Puro (Addgene #236081...breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked by unique restriction sites...plasmid set, our bacterial plasmid set for expressing tagged proteins, and stay tuned for mammalian expression...streptavidin immunoprecipitation (IP), and tandem mass tag (TMT) mass spectrometry (MS) in HCT116 cells (colorectal...by the Loren Looger and Kaspar Podgorski Labs now has a new version: iGluSnFR4, developed by the GENIE ...we're excited to share our very own set of empty, HA-, and EGFP-expressing lentiviral backbones designed...

...Advanced Microscopy (University of Amsterdam). Tagging a protein of interest with a fluorescent protein...still block the function or activity of the tagged protein because of its size. This type of interference... is known as “stickiness.” Since the fluorescent tag should operate as an inert light emitting module,...successful insertion of a fluorescent protein are the tagging of Ccd42 (Bendezú et al., 2015) and the alpha subunit...most notably CRISPR/Cas mediated HDR) enables the tagging of endogenous genes close to native expression ...protein variant, and where to add the fluorescent tag. These choices will determine how well the fusion...reflects the function of the native protein that is tagged. The localization of the fusion protein can be ...

...extracts (Gagoski et al 2015). He has also constructed a library of eGFP-tagged human ORF clones to allow testing...-terminal eGFP tag carried on all constructs. The Cell-Free Protein Expression Kit has been used to produced...additionally contain various combinations of protein tags (His, Avi, and GFP) as well as protease sites to...researcher to quickly generate N- and C-terminal epitope tag fusions to a protein of interest. The plasmids used...folding and/or protease sites to remove the epitope tags. Different plasmids in the kit can be used to purify...specific protein folding needs. The pCri system has recently been used to study a matrix metalloproteinase... Queensland’s Institute for Molecular Bioscience has provided a solution for this problem by developing...

...pAAV-TK-Acceptor has a conventional heterologous promoter-driven TK-neoR gene. To modify these tagging vectors ...their lab include using AAV to introduce epitope tags into the endogenous alleles of the p53 and PTEN ...allele of the STAG2 tumor suppressor in human cells (Kim et al, 2016). The Waldman lab has designed their...their AAV tagging vectors (pAAV-SEPT-Acceptor and pAAV-TK-Acceptor) so that they can be easily adapted ... introduction of heterozygous endogenous epitope tags, or the introduction of dominant mutations of a ...: PMC373311. 3. Kim, Jung-Sik, et al. "Epitope tagging of endogenous genes in diverse human cell lines... at the Georgetown University School of Medicine has used this gene editing system frequently over the...

...Tet-on transgene vector. Degron tags Tagging relevant proteins with a degron tag can provide inducible protein...systems work by tagging the endogenously or exogenously expressed protein with the relevant tag and then introducing...ligand to mark the tag and whatever it is fused to for degradation. Since the tagged protein of interest... are akin to degron tags, just with a flipped mechanism. When a protein is tagged with a DD, it is unstable...auxin-inducible degron (AID), and degradation tag (dTAG). These control systems function at the post-translational...Check out Addgene's Lentiviral dTAG cloning vector and SMASh degron tagging vector! Destabilizing domains...quickly recover once the ligand is removed. Degron tag systems include small molecule assisted shut off ...

... well as additional useful tags, including 3xFLAG epitopes, TEV and BioTag, which can be used in protein... protein effectivity), and a fluorescent protein tag for visualization (Fig. 1b). While the probes target...ultimately leading to the degradation of the AID-tagged protein. To quantify protein degradation the Ward...44 amino acids auxin inducible degron sequence) tagged proteins. Degradation of your protein interest ...Gibson and Saptrap cloning systems to generate AID* tagged CRISPR knock-in repair constructs. These repair...repair constructs can be used to integrate the tagged AID* into genomic sites of interests via CRISPR techniques...Cecchetelli The auxin inducible degron (AID) system has been extensively used in C. elegans since 2015 to...

...superfolder GFP (sfGFP) with the small solubility tag GB1. In SunTag nomenclature throughout the rest of this ...tracking Caveats of SunTag Very large size: a 24x scaffold fully occupied with GFP has a molecular weight...References 1. Tanenbaum, Marvin E., et al. “A protein-tagging system for signal amplification in gene expression...concerns are greater with SunTag. v1 SunTag exhibits some scaffold aggregation (v4 Suntag does not.) It’s up...may be toxic to the cell. Here comes the SunTag How does SunTag fix these problems? Instead of directly...power of SunTag for single molecule imaging, finding that plasma membrane-targeted CAAX-SunTag was 18-fold...traditional FP fusion, a SunTag fusion could be used as well. Pros of SunTag compared to traditional ...