We narrowed to 95 results for: Myc-tagged Cre

...electron microscopy (EM) tag providing superior subcellular localization of tagged proteins compared to traditional...additionally incorporates SUMO tags in the FP sequence for separation from Cre-based Brainbow-expressing cells...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...of an isopeptide bond between 2 peptide tags, SpyTag and KTag. The group demonstrated the use of the SpyLigase... short hairpin) and CRUSH (Conditional RUSH) use Cre-mediated recombination to turn on or off the expression...a dual-color RNAi “sensor” mouse strain in which Cre expression causes the induction of both DsRed fluorescence...

... a unique tag into a genomic site of choice via the mutational abilities of PolI3m. This tag would allow...modular nature, the Dueber Lab created a few versions of EvolvR. To increase the mutation rate of EvolvR,...with a targeted mutagenesis rate ~9-fold higher than the original nCas9, while increasing the standard mutation...mutations to PolI3M generated PolI5M which increased EvolvR’s mutagenesis rate to ~10-3 mutations per nucleotide...bacteriophage T7 DNA polymerase increased the processivity of PolI3M, and thereby increasing the editing window of...result in a spectinomycin resistant phenotype in bacteria. While there are many known spectinomycin resistance... for mutagenesis, a feature which they used to generate bacteria resistant to both spectinomycin and streptomycin...

...construct, the neomycin resistance gene is floxed so that it can be removed post-selection through Cre-mediated...want to create a conditional knockout. To make a conditional knockout, researchers often use the Cre/Lox ...researchers usually remove it after selection using the Cre/Lox recombination system. After “floxing” the resistance... you can remove the gene through the addition of Cre recombinase (Figure 2). Figure 2: A knockout...make it possible to insert, or knock-in, any gene, tag, or mutated exon into the genome. For this purpose...example, you can remove a floxed resistance gene using Cre recombinase. If GFP is under control of an endogenous...promoter responds. You can also use this method to tag an endogenous protein with GFP, as seen in blue flame...

...protein tags can be used to facilitate the visualization of specific proteins in cells. Tagged protein...containing AHA to a fluorescent alkyne tag. The newly synthesized and tagged proteins can then be observed with...the knock-in of fluorescent tags into the gene locus of interest. An advantage of this alternative is the...here, there is the question of whether or not the tags influence behavior. A conference and a breakthrough... known as fluorescence noncanonical amino acid tagging (FUNCAT, Dieterich et al. Nature Neuroscience, ...protein of interest that also had incorporated the AHA tag after pulse labeling of a determined period of time...used for cell labeling Check out other protein tagging strategies Additional Resources on Addgene.org...

...expression ~ ~ ~ ~ ✓   ✓ SV40 and c-Myc nuclear localization sequences Improve translocation..., the Liu lab created and evaluated 19 PE1 variants with RT mutations known to increase activity, enhance... initial nick site (to avoid creating a double strand break), increasing editing efficiencies 2-3-fold... Figure 1: An overview of prime editing. Created with BioRender.com. How does it work? First...ones in Addgene’s repository later in the post. Advantages of prime editing Less constrained by PAM sequence... between the template and primer binding site, increase processivity, or improve thermostability. What...fold. However, the process of double nicking increases indel formation slightly. Designing the sgRNA with...

... SpCas9 is also fused to a 3X FLAG tag at its N-terminus. This tag can be used to test for SpCas9 expression...Alternative forms of this vector contain GFP-tagged and un-tagged versions of SpCas9 and further variants ...SpCas9 is fused to the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables...causes the ribosome to translate Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This...predecessor, which had a point mutation in the puromycin resistance gene that made it less effective in...SpCas9n (nickase), which nicks DNA rather than creating double stand breaks and may better facilitate ...

...found in Botman et al., 2019 . FP Tagging Yeast tagging vectors to create fusions of proteins of interest...Optimized cassettes for fluorescent protein tagging in Saccharomyces cerevisiae. Yeast, 21 (8):661-70. https...plasmids to overexpress various fluorescent markers and tag your proteins of interest with fluorescent proteins...Collection Botman-Teusink Yeast FP Collection FP Tagging FP Overexpression Resources This comprehensive ...allows for constitutive overexpression of FPs and tagging of genes of interest with FPs in yeast. Characterization...TCGATGAATTCGAGCTCG–3' ID Plasmid Selectable Marker Tags Publication FP Overexpression Plasmids for constitutive...Improved blue, green, and red fluorescent protein tagging vectors for S. cerevisiae. PLoS One, 8 (7):e67902...

... pSico 3rd Conditional (Cre-lox), stable expression of shRNAs; addition of Cre turns on shRNA expression...102930 pLJC5-Tmem192-3xHA 3rd Contains a lysosomal tag. See Abu-Remaileh et al. (2017) for additional variants...shRNA empty plasmid with puromycin selection. See plasmid 24150 for hygromycin selection. Bob Weinberg ...expression of shRNA with puromycin selection. See plasmid 21916 for neomycin selection. Dmitri Wiederschain...expression of shRNA with neomycin selection. See plasmid 21915 for puromycin selection. Dmitri Wiederschain...19319 pLJM1-EGFP 3rd CMV-driven EGFP fusion with puromycin resistance. Can be used for cDNA expression. David...pLenti-puro 3rd CMV-driven expression of cDNA with puromycin selection. Ie-Ming Shih 25737 pSLIK-Hygro 3rd ...

..., modular plasmids, combining fluorescent tags, epitope tags, protease sites, and selection markers. These...fluorophore tagging vectors for your single or multi-color imaging experiments. Interested in epitope tags? Others...have genome tagging systems enabled you to advance your research? Do you have a favorite tagging system that...Researchers have long taken advantage of this natural process to integrate protein tags into the genomes of S...multiple cloning site in place of the epitope tag for creation of your own unique protein fusions. This collection...promoter substitution), and tagging at either the N- or C-terminus (3xHA, 13xMyc, GST, or GFP). Longtine ...especially useful in protein complex studies where tagging of multiple protein products is desired, as multiple...

...transgene expression. Which tag should I use? When deciding on the appropriate tag, the experimental design...our studies suggest that non-fluorescent tags may be advantageous over fluorescent proteins when the fluorescence... methods, these proteins are routinely fused to ‘tag’ or ‘label’ proteins to facilitate its detection....detection. Fluorescent proteins are most widely used tags because they emit endogenous fluorescence that allow...protein (GFP) and mCherry, two of the most common tags currently used, are approximately 240 amino acids...most of them were still cytoplasmic. Use of ‘small’ tags to identify proteins Given the limited expression...monkey neurons, we next hypothesized that a different tag fused to the hM4Di might improve the transport of...

...cloned into an IPTG-inducible His-tag expression vector. All His-tags are cloned at the C-terminus of ...inexpensive to assay, and easy to purify using histidine-tagged constructs and routine protein and molecular biology...a TEV cleavage site is present between the 6xHis-tag and the coding region of MDH. For detailed clone ...Organism or species (e.g., human, watermelon, Streptomyces ) Subcellular compartment (chloroplastic, cytoplasmic.... PMID: 36893201 (Link opens in a new window) Increasing access for biochemistry research in undergraduate...

... 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...engineering plasmid tools in about a week. Taking advantage of Golden Gate cloning, the Haynes lab’s Protocols.io...

...KB Fujii iChIP/enChIP to purify genomic DNA FLAG tagged dCas9 PDF, 110 KB Goldstein Nematode: gRNA design...pCFJ90 (pharyngeal co-injection marker); pDD104 (Cre recombinase) PDF, 105 KB Jaenisch CRISPR plasmid ...PDF, 109 KB Mendenhall and Myers Mammalian: FLAG tagging endogenous proteins pFETCh_Donor ; additional HDR...- a CRISPR Screen Tool (Link opens in a new window) Design gRNAs for custom CRISPR screens targeting a...in your lab, from knockouts to pooled library screening. CRISPR eBook Addgene's downloadable eBook about...CRISPR pooled libraries for large-scale functional screens. Available for knockout, activation, inhibition...For use with genome-scale CRISPR-Cas9 knockout screens (or GeCKO) technology. Developed by the Xiaole ...

... (CAPTURE.) Tag : Multiple methods make it easier to tag endogenous loci with epitope tags or fluorescent...protein tags, or loxP/FRT sites, among others. Homology-directed repair (HDR) allows us to create a variety... not occur in mammalian cells. Purify : Epitope-tagged dCas9 can also be used to purify a genomic locus... EM, Myers RM. 2015. CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res . ...with work directed towards increasing on-target editing rates and decreasing off-target editing rates. ... locus makes it especially amenable to genetic screens. Pooled gRNA libraries can be used to identify ...sequencing, CRISPR is a robust system for genome-wide screening. CRISPR Technological Advances CRISPR technology...

...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...2015). He has also constructed a library of eGFP-tagged human ORF clones to allow testing and comparison...conveniently performed through the N-terminal eGFP tag carried on all constructs. The Cell-Free Protein ...greatly reduced costs per plasmid. We generally create kits to allow you to easily use a set of plasmids...markers built into all plasmids in the kit help decrease false positives observed in the cloning process...

...The permanent labelling of cells using cre-lox is advantageous when compared to photoactivable or photoconvertible...cells according to whether or not they have been tagged with a fluorescent protein or dye. It separates... distinguishing cell populations that have been tagged with different fluorescent proteins or dyes. 3....useful tool for marking and tracking cells is the the Cre/lox recombination system. A researcher can flank ...expression is turned on or off in cells expressing Cre. The result of a system like this is a mosaic of ...progenitor cell migrating to the epithelium). At this stage, only cells from the immune system show wide scale... In the past 15 years efforts have been made to create transgenic mice expressing fluorescent protein ...

...Plasmid Collections Recombinases Cre-Lox and Other Site-Specific Recombinases Cre Plasmids Flp Plasmids Dre ...prevent gene expression in the absence of Cre. In the presence of Cre, the stop codon is excised, and gene ...until Cre is present, at which time the gene will be disrupted or deleted. shRNA Expression: Cre-lox can...interest, Genes A and B. When Cre is absent, only Gene A is translated correctly. Cre expression excises Gene...double-crossover event. Toxicity Cre-lox recombination is a very useful tool, but Cre expression can be toxic ... such as Cre-only lines to account for cellular toxicity in your system. Using inducible Cre systems in...expressing Cre and other site-specific recombinases. Many of our in-stock viral preps feature Cre-dependent...

...terminally tagged fusion proteins with fluorescent (YFP, GFP, CFP, RFP, etc.) or affinity tags (HA, FLAG... plant selection markers (basta, hygromycin, kananamycin, tunicamycin) (Nakagawa et al., 2007). Alternatively...new inflorescences begin to grow. This will also increase the number of inflorescences overall, which may...them again.  This won’t hurt them, and it will increase the chances of tDNA integration. Selecting Arabidopsis... seeds too densely during selection: this will increase background growth of non-transformants and make...Gateway Binary Vectors: High-Performance Vectors for Creation of Fusion Constructs in Transgenic Analysis of...

...tissue-specific cell types: Cre-lox recombination. What is Cre-lox? The Cre-lox system is a technology ...marker, Cre can be used to easily perform this eviction. Regulated Cre expression – placing Cre downstream...constructed. Additionally, Cre-containing adenovirus (Ad-Cre) or AAV (AAV-pgk-Cre) has been used to successfully...generated at the loxP sites. How can I use Cre-lox? The Cre/lox system is a well-established research ...prevent gene expression in the absence of Cre. In the presence of Cre, the stop codon is excised, and gene ...facilitate the use of Cre-lox technology, transgenic mice have been constructed that express Cre under a variety...analogous to Cre and loxP, respectively. The FLP-FRT technology can be an effective alternative to Cre-lox, and...

...Vectors Read our Blog Post About Tagging Yeast Genes ... Scientists took advantage of these discoveries to create a new system for screening protein-protein interactions... developed in Saccharomyces cerevisiae in 1989 and are still used extensively to screen for molecular ...identifying prey proteins after screening. It is also possible to screen libraries of bait and prey against...single-pass two-hybrid screening (6). See the Tips section for more on how to increase your rate of interactions...interactions when screening! Tips for two-hybrid screening Minimizing false positives: Run replicates of ...protein interact and enable intein splicing (5). Screening with yeast two hybrid systems While initial studies...