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We narrowed to 260 results for: gfp gene

Showing: 161 - 180 of 260 results

  1. Hot Plasmids February 2024

    Type
    Blog Post
    ...-invasive systemic gene delivery in non-human primate (NHP) brains.  Systemic gene delivery capsids previously...Monomeric StayGold variants AAV.CAP-Mac for systemic gene transfer in primate brains Endothelial-specific ..., Myc and SV40 Nuclear Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral...Adeno-associated viral vectors for functional intravenous gene transfer throughout the non-human primate brain....plasmid here! Chen, X., et al. (2023). Functional gene delivery to and across brain vasculature of systemic...past decade, primary cells remain difficult to genetically edit due to toxicity from electroporation or ... under CC-BY license.   The Miyawaki lab generated a series of StayGold mutants and identified a ...

  2. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...contain a toxin gene (regulated by a rhamnose-inducible promoter) and a chromoprotein gene. Allelic exchange...contrast, the widely used green fluorescent protein (GFP) gives a bright green fluorescence when exposed to...the plate. For example, if your plasmid contains a gene encoding a blue chromoprotein, after transforming...3: pTOX plasmids expressing amilCP and tsPurple genes. Image: Lazarus et al., 2019. One such lab is...protein family. What sets them apart is the way they generate their color: they strongly absorb visible light...difficult to directly use in bacteria, where most genetic manipulations start. Anthony Forster’s lab used...eforRed, asPink, spisPink, scOrange, fwYellow, amilGFP, amajLime, cjBlue, meffBlue, aeBlue, amilCP, tsPurple...

  3. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ...under-expressing clones. If GFP-encoding cDNA is co-transfected at a favorable ratio to the gene of interest (or,...overexpress genes of interest; more recently we have also been using Addgene CRISPR vectors to generate cell ...cell lines with knockouts of specific genes. Many years ago, people in the laboratory became frustrated with...with different levels of expression of our gene of interest. Advantages of the agarose cloning method ...figure 1.   Find Additional Useful Protocols on Addgene's Protocol Pages Agarose cloning method procedure...-well cell plate to a 96-well assay plate. We generally freeze at least 2 ampules each of 4 clones with...som.umaryland.edu.   Additional Resources on the Addgene Blog Tips for a 1st Time CRISPR User (by a 1st...

  4. Zhang Lab CRISPR Page

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    Collection
    ...activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator helper complex with 2A GFP 61424 : sgRNA...(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). This is a AAV plasmid for sgRNA cloning. GFP-KASH fusion ... information, please visit the Addgene SAM library page . Targeted gene activation using SAM There are...available for mammalian endogenous gene activation using SAM: Addgene plasmids #61422 - #61424 were designed...contain a combination of CRISPR-associated (Cas) genes as well as non-coding RNA elements capable of programming...for the transcriptional activation of endogenous genes. It consists of three components: A nucleolytically...human codon-optimized, and packages both the SaCas9 gene and its single guide RNA (sgRNA) into one plasmid...

  5. Serotype Testing AAV

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    Collection
    ...AAV1). AAV Vectors for Serotype Testing pAAV-CAG-GFP (Plasmid #37825) Description : Ready-to-use AAV in...plasmid 37825 (deposited by Edward Boyden ) and direct GFP expression from the CAG promoter. For information...Ready-to-use AAV available from Addgene's viral service. Control EGFP vectors in various serotypes for... 37825-AAVrg.T 20 µL $ 150 Add to Cart pAAV-hSyn-EGFP (Plasmid #50465) Description : Ready-to-use AAV ...plasmid 50465 (deposited by Bryan Roth ) and direct EGFP expression from the human synpasin promoter. For...

  6. Trimmer Lab NeuroMab Collection

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    Collection
    ...199419 Anti-GFP [N86/38R-2b] GFP Aequorea victoria Mouse IgG2b 199420 Anti-GFP [N86/8R-2b] GFP Aequorea ...206719 Anti-GFP [N86/38R-1] GFP Aequorea victoria Mouse IgG2a 206720 Anti-GFP [N86/8R-1] GFP Aequorea victoria...Rat Mouse 206743 GFP scFv [N86/44] N86/44 scFv GFP Aequorea victoria Mouse 206744 GFP scFv [N86/20] N86...short and long Rat Mouse IgG2a 114492 Anti-GFP [N86/38.1R] GFP Aequorea victoria Mouse IgG2a 114493 Anti-NGL...glutamate receptor Rat Mouse IgG2a 177572 Anti-GFP [N86/8R] GFP Aequorea victoria Mouse IgG2a 177573 Anti-...N479/107R] VAPA/B Mouse IgG2a 188164 Anti-GFP [N86/44R] GFP Aequorea victoria Mouse IgG2a 188165 Anti-.../22R] Prrt2 Mouse Mouse IgG2a 188166 Anti-GFP [N86/20R] GFP Aequorea victoria Mouse IgG2a 188167 Anti-...

  7. Plasmids 101: The Promoter Region – Let's Go!

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    Blog Post
    ...Vectors Read about Reporter Gene like Luciferase and GFP Resources on Addgene.org Find Plasmids for Your...depending on its orientation to your gene. T7lac High levels of gene expression Promoter from T7 bacteriophage...modulating gene expression through varied inducer concentrations. trp High levels of gene expression...human or mouse) General expression mRNA Mammalian promoter from phosphoglycerate kinase gene.  Constitutive...deacetylation. Ubc General expression mRNA Mammalian promoter from the human ubiquitin C gene Constitutive  ... human beta actin General expression mRNA Mammalian promoter from beta actin gene Constitutive  Ubiquitous... Requires the presence of Gal4 gene to activate promoter. Ac5 General expression mRNA Strong insect ...

  8. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...sequence (UAS) of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether...promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...reporter gene (3, 4), there are now many variations, including many that use auxotrophic rescue genes as reporters...Shapira S.K., Chou J. Beta-galactosidase gene fusions for analyzing gene expression in Escherichia coli and ...hybrid system (also called the interaction trap), genes encoding proteins of interest(POI’s) are fused to... in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing the target... the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI’s interact...

  9. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...the ccdB gene is replaced by the gene of interest. As a result, cells that retain the ccdB gene die off...tool. The plasmid pKSCTMF contains the mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR...The team used this cassette to knock out genes, knock in a GFP cassette, and introduced site-directed mutations...antisense RNA regulates hok gene expression indirectly through the overlapping mok gene." Journal of molecular...associated genes. These systems better ensure that plasmids are passed down through generations (Figure ...application of CcdB is GatewayTM cloning, where the ccdB gene serves as a counterselectable marker in the destination...toxin-antitoxin system to construct a stable synthetic gene circuit that can be used over prolonged periods....

  10. Tips for Using FRET in Your Experiments

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    Blog Post
    ...Express a gene of interest fused to the C-terminus of SCFP3A Amber N1 Yellow Mammalian Express a gene of interest...Express a gene of interest fused to the C-terminus of Amber mVenus N1 Yellow Mammalian Express a gene of interest...Clover (a GFP variant) commonly used with mRuby2 pLSSmOrange-N1 Orange Mammalian Express a gene of interest...your choice of fluorescent fusion protein with your gene of interest: Plasmid Color Expression Description...Aquamarine mCerulean N1 Cyan Mammalian Express a gene of interest fused to the N-terminus of monomeric...Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus of monomeric...monomeric Venus mVenus C1 Yellow Mammalian Express a gene of interest fused to the C-terminus of monomeric...

  11. Fluorescent Proteins 101: Fluorescent Protein Timers

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    Blog Post
    ...Resources on the Addgene Blog Learn More About Fluorescent Protein Aggregation Catch Up On Your GFP Basics Get...expression time. 2. Timer functionality with a single gene Although it is possible to make a timer system composed...cargoes to FP timer mk-GO (Tsuboi et al.) Monitor gene expression in the developing pancreas by placing...FP timer DsRed-E5 under the control of Neurog3 (a gene controlling pancreatic differentiation) in mouse...(trafficking to a subcellular location, start of gene expression, development of a cell structure, etc...formation of limbs are correlated with changes in gene expression. In a developing embryo, for example,...long-lived transcription factors control gross changes in gene expression as new body sections are formed. In synthetic...

  12. Simplify Cloning with in vivo Assembly

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    Blog Post
    ..., or a mutation can be introduced in a gene, while adding a GFP-tag (Figure 3). Achieving multiple modifications...insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer design for each type...amplified, restriction digested or even a synthesized gene. PCR is the primary method to produce the linear...included in a single PCR, for example for subcloning genes from different templates. All PCRs are run as an... 5’ end of either primer. Subcloning - To move a gene from one vector to another simply amplify the vector... to the 5’ end of the reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride...using the same three step protocol:  Fragment generation - Any linear DNA fragments can be assembled in...

  13. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ... a superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage... (Shmakov et al., 2015). They were using Cas1, a gene commonly associated with CRISPR arrays and involved...analysis, they identified 53 potential candidate genes that fell into 3 categories based on the architecture...expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although the...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Check Out Our CRISPR Featured...27, 2020. CRISPR, and specifically Cas9 from S. pyogenes (SpCas9), is truly an exceptional genome engineering... PMC5127784. Find plasmids from this paper at Addgene. 2. East-Seletsky, Alexandra, et al. "Two distinct...

  14. Tips for arabidopsis transformation

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    Blog Post
    ...swapping and generating N and C terminally tagged fusion proteins with fluorescent (YFP, GFP, CFP, RFP, ...transcriptional and translational dynamics of genes and proteins in living plants, to complementing mutant phenotypes...potato contains Agrobacterium T-DNAs with expressed genes: An example of a naturally transgenic food crop....simplifying plant genetic engineering Using arabidopsis in the classroom Resources on Addgene.org Find plant... transformation. There are many motivations to generate transgenic Arabidopsis, from studying transcriptional... floral drip or dip transformation method. The general steps for this method include: Cloning and transforming...Glazebrook, 2002). Find plant plasmid resources at Addgene  Choosing an expression system Choosing an appropriate...

  15. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...negative bacteria periplasm. Efforts to tag endogenous genes for proteins in these compartments using CRISPR ...tagging to detect proteins expressed from cloned genes: deletion mapping functional domains of Drosophila...Lindsey M. Costantini, and Erik L. Snapp. "Superfolder GFP is fluorescent in oxidizing environments when targeted...quantitative imaging experiments and cell physiology in general. Pitfalls of Fluorescent Protein Fusions Observing...cellular compartments other than the cytoplasm. Generally, FPs evolved in or were engineered for use in ...N-glycosylation consensus sequences in popular FPs. This generated a palette of inert green, cyan, yellow, and blue...standard FPs. The moxFPs are now available through Addgene Many thanks to our guest bloggers Erik L. Snapp...

  16. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...check out our article: Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection. Cellular...been generated for yeast (Kim et al., 2019) and fruit flies (Bischof et al., 2018), the generation of such...quantities (and the correct stoichiometric ratios) to generate useable data. What’s more, measuring and quantifying...straightforward nature of BiFC also makes it easier to generate a large number of constructs for high-throughput...screening of PPIs. For example, Bischof et al. (2018) generated an open reading frame (ORF) library to screen ... such resources in less genetically tractable organisms lags behind. BiFC can also be used to screen for...pair. Find BiFC plasmids for your experiment at Addgene! How to Design a BiFC Experiment Much like FRET...

  17. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...which must be stabilized by monomeric superfolded GFP and averages only ~50% RNA knockdown. In initial ...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Check out our CRISPR Featured...and designated this system REPAIRv1. Using next-generation sequencing, they confirmed A->I editing and found...cytosine deaminase. Three rounds of rational mutagenesis on the ADAR2DD residues that contact the RNA ...RNA-sequencing, the authors performed rational mutagenesis and identified that a S375A mutation in RESCUE...PMID: 29070703. Find plasmids from this paper at Addgene. Abudayyeh, Omar O., et al. “RNA targeting with...PMID: 28976959. Find plasmids from this paper at Addgene. Abudayyeh, Omar O., et al. "C2c2 is a single-...

  18. Antibodies 101: Immunoprecipitation

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    Blog Post
    ...In cases such as this, an epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest ...resources on the Addgene blog: Introduction to Immunofluorescence View all of Addgene's antibody posts  ...Resources on Addgene.org: Find plasmids encoding recombinant antibodies at Addgene  ...that interact with nucleic acids. Labs studying epigenetics routinely use the chromatin IP (ChIP) technique... found this blog helpful be sure to check out Addgene’s blog for other antibody topics.     References...

  19. The Challenges of Cell Culture

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    Blog Post
    ...about the Dangers of Using GFP for Protein Localization Resources on Addgene.org Find Plasmids for Mammalian...pharmacology (many of which are represented in Addgene's Special Collections). Arguably, oncology has benefited...examining whether the cell lines had isoenzymes and genetic polymorphisms specific to their origins. This technique...out by the commercial repository. Repositories generally ensure cell line identity and purity making it...to the original as possible and so any results generated are reproducible and reliable. When you order ...Abcam Cell Culture Guidelines Resources on the Addgene Blog Learn All about Mammalian Vectors Read up...

  20. New England Biolabs Cell-Imaging Plasmid Collection

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    ...comprehensive comparison to GFP, please refer to NEB's comparison of SNAP-tag, CLIP-tag, and GFP . Technology SNAP-tag...England Biolabs has deposited plasmids at Addgene to generate SNAP, CLIP, ACP or MCP tagged plasmids for...associated with this technology are available at Addgene for distribution to the research community. Key...synthases needed for use of these tags. Plasmids Addgene ID Plasmid Substrate Feature 101137 pSNAP-tag (...and the ACP and SFP Synthases. Users may still generate their own fluorescent or non-fluorescent CoA substrates...
Showing: 161 - 180 of 260 results