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We narrowed to 407 results for: expression vector

Showing: 201 - 220 of 407 results

  1. Antibodies 101: Affinity Tags

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    Blog Post
    ... single protein is expressed, with the intent of purification, it will be expressed along with all other...other cellular proteins within the expression system (bacteria, yeast, mammalian, etc.). To individually...about that protein that no other protein in the expression system has. This is where affinity tags come ...peptides which do not naturally exist in most expression systems that can be added to the N or C terminus...on: The protein you are trying to purify The expression system If you want the tag to remain on your ...MBP tag, although large, can actually increase expression and solubility of the tagged protein (Fox and...Ready to start tagging? We are here to help with vectors, protocols, antibodies, and more!   Resources ...

  2. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

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    Blog Post
    ... information on viral vectors, including transduction download our Viral Vectors 101 eBook. Bacterial ...bacterial or eukaryotic cells via a virus or viral vector. One example are bacteriophages that attach to ...scientists commonly use phagemids, a DNA cloning vector that contains both bacteriophage and plasmid properties...transient cell lines, where a gene of interest is expressed but not integrated into the genome and stable ...

  3. Antibodies 101: Designing Your First Flow Panel

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    Blog Post
    ...transgenic protein for the HEK 293T cells to express: CD45. Upon expression, CD45 is incorporated into the plasma...CD45, but is separately expressed. (The reason for having a separately expressed transduction reporter ...manage to express CD45. Thus, if transduction efficiency is much higher than CD45 expression, this could... example, point at issues with the double-expression vector.) In our flow experiment, we'll want to identify...CD45+ cells (CD45 expression) and assess our transduction efficiency (eGFP expression).     The first ...detected through a cell surface label. The viral vector also encodes for eGFP, which serves as a transduction...rest of our panel. Lastly, we want to detect the expression of our protein of interest: CD45. I am using ...

  4. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

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    Blog Post
    ...genome editing, activating gene expression, repressing gene expression, and much more. But there’s one... T, Lieber A (2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR/Cas9... means they can be delivered in vivo using AAV vectors. Having a way to shut off CRISPR-Cas systems prevents...

  5. Plasmids 101: Inducible Promoters

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    Blog Post
    ...inducible system is found in many bacterial expression vectors. Negative inducible promoter pBad is another...gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant...Temperature inducible promoters Temperature sensitive expression systems are typically less leaky than chemically...chemically induced promoters; they show near-zero expression at regular temperatures but can be induced by ... promoters, in which a gene of choice is only expressed following exposure to a brief heat shock. In the...promoters Light is another way to activate gene expression, and two-component systems used in synthetic ...transcription from phage promoter pR, preventing expression of a reporter gene. When light is present, YFI...

  6. Custom CRISPR Screens & the Green Listed Software

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    Blog Post
    ...screens Differentially expressed genes  As the costs for global gene expression analyses like RNAseq are...test which of the differentially expressed genes found in a gene expression dataset are causal for a phenotype...differentially expressed genes that are known drug targets, or all differentially expressed genes that code...sequencing to identify which gRNAs are differentially expressed in the separate groups and, as a consequence, ...CRISPR screens designed to target differentially expressed genes can be the next stage in making sense of...interactions to a large extent involve receptors expressed on the cell surface. Thus, a screen can be used...The gRNA oligos are subsequently cloned into your vector of choice using the adapter sequences that you ...

  7. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

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    Blog Post
    ...genes is from Proteobacteria to mammalian expression vectors. And more broadly, their data revealed that...a distantly related organism, the gene may not express well because of these different codon usage biases...closely resembles that of the host organism, it may express protein more efficiently. To classify gene sequences...rare codons, sometimes intentionally modulating expression in ways we don’t quite understand yet,” Kunjapur...synthetic. They also found out that the most common expression system within the Addgene repository is mammalian...

  8. Plasmids 101: Restriction Cloning

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    Blog Post
    ...instance, if you were cloning a gene into an expression vector, you would want the start of the gene to end...Your Insert and Vector by Gel Purification     Now that you’ve cut your insert and vector, unfortunately...unsuccessful 1), in a vector alone plate with many more colonies than the vector + insert plate (unsuccessful... ligation step. 3. Ligate Your Insert into Your Vector In the ligation step, you mix your purified, cut...Resources on Addgene.org Find the Perfect Empty Vector for Your Cloning Experiments Browse Our Molecular...

  9. Another Pathway into Cells: iTOP

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    Blog Post
    ... mutations and gene expression changes elsewhere in the genome. Second, expression is by nature transient...control could enter cultured cells and activate expression of a luciferase reporter. CPP was not required...stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. CRISPR...causes a cell to take up, and continued protein expression would require multiple transduction events. For... to binary systems in which transient protein expression has a long-lasting effect, such as Cre- or Cas9...limitations of packaging size associated with viral vectors. For these reasons, iTOP, especially in tandem ...

  10. Hot Plasmids - November 2023

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    Blog Post
    ...fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4 from different promoters...to reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro applications...activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive element...to create a three-dimensional atlas of protein expression in the adult mouse brain using thick-section ...

  11. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

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    Blog Post
    ...interest. In our studies, we used AAV viral vectors to express hM4Di in neurons of rhesus monkeys or mice...successful expression of the protein of interest, an alternative could be bicistronic expression of the fluorescent...the fluorescence expressed in the neuropil conceals identification of opsin-expressing cells). For other...proteins provide a versatile method to identify the expression of transgenes of interest. Furthermore, fluorophore...can be a roadblock, rather than an aid, in the expression of the protein of interest. The green fluorescent...’ tags to identify proteins Given the limited expression of hM4Di‐mCherry at the plasma membrane in monkey... mCherry with the HA tag resulted in abundant expression of hM4Di in plasma membranes in both species ...

  12. CRISPRainbow and Genome Visualization

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    Blog Post
    ...can be expressed in a single vector, pCRISPRainbow-DONOR1, with dCas9 supplied by a separate vector. ...CRISPRainbow gRNA will permit labeling, but a newly expressed standard length gRNA would “switch” the system...

  13. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

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    Blog Post
    ...years, he and his colleagues built a Molecular Expressions collection including some 3,300 plasmids along... Baird said. “They are all designed in the same vector backbone with very similar restriction enzymes....shutter its doors. “We wanted to give all of these vectors to the scientific community and contribute in a...

  14. Developing a cold tolerant E. coli using specialized chaperones

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    Blog Post
    ... compared to the E. coli strain expressing the empty vector. And thus was born ELSA - our happy lil’ cold-tolerant...decided to focus on a special class of enzymes expressed by psychrophilic (cold-loving) bacteria. These...recombinant psychrophilic enzymes for the host The expression of recombinant proteins for industrial or research...mesophilic, for example E. coli, carrying a plasmid expressing a protein of interest). Mesophilic microbes are...enzymes face very similar issues when forced to express in mesophilic host systems. The so-called “regular... to grow at low temperatures suitable for the expression of psychrophilic proteins. The primary problem...Factor), that would allow the stable folding and expression of recombinant psychrophilic proteins. As the...

  15. Celebrate Open Data Day with Addgene’s AAV Data Hub!

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    Blog Post
    ...methods did you use for expression of AAV cargo? Did you use GCaMP expression? Direct Fluorescence? Immunohistochemistry... visualized expression after AAV injection, what region and cell class ended up expressing your virus?...Addgene.org Read our AAV guide Browse all  AAV vectors Find ready-to-use AAV preps ...

  16. Mouse Modeling, Part 2: Breeding and Crossing Mice

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    Blog Post
    ... that expresses Cre with a strain that expresses a floxed gene of interest. Cre may be expressed globally...the appropriate primers to detect your targeting vector or gene of interest. 2. Breed your parental mice...infertility and can arise due to copy number, expression level, and site of integration of the transgene...in cell lines to determine how it impacts the expression level of the transgene. In existing strains of...heterozygous background to reduce copy number or expression level effects.  Another challenge that may arise...heterozygous for each mutated allele.  Cre-lox expressing strains Cre-lox strains are a common example ...we explained how genetic recombination in mice expressing both Cre and LoxP occurs. But how are these mice...

  17. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

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    Blog Post
    ...working on developing new surface display vectors to improve expression and trafficking of these biosensors....gray (PAG), with red neurons expressing tdTomato and green neurons expressing cFos. Image from Elizabeth... Figure 2: Micrographs of the biosensor expressed in the human cell line, Hep3B. From left to right...opioid withdrawal with tdTomato and assessed cFos expression. She will explore the connectivity patterns of...chemogenetics to study the activity of parvalbumin (PV)-expressing neurons, neurons that the lab previously found...the receptors to monitor their localization and expression. Qiu saw that inhibition of the PV neurons resulted...response due to blue light stimulation of dual virus-expressing CR neurons. (C) Duration of painful responses...

  18. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

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    Blog Post
    ...then easy to package in low-capacity vectors like AAV, a popular vector due to its low immunogenicity. But...in size and are easy to package in low-capacity vectors! These small, but mighty type VI-D enzymes are ...collateral cleavage does not occur when Cas13d is expressed in a mammalian system. Since Cas13d is functionally...

  19. Common Injection Routes in Mice

    Type
    Blog Post
    ...be injected, including cells, drugs, DNA, viral vectors, and viruses. All researchers must go through ...choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal injections...for the delivery of various therapies and viral vectors. This injection sounds very disruptive, but surprisingly... This is most often used for delivery of viral vectors, though drug treatments are also possible. One ...Addgene.org Genome Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox plasmids References Prathipati...

  20. Hot Plasmids - February 2022

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    Blog Post
    ...individually expressed proteins run in separate lanes. B: individually expressed and coexpressed proteins ...through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either 10, 30, 50...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure... Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes when encoding multiple guides. While...opportunity for lab members or students new to protein expression and purification. Figure 3: The Penn State...Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60, 80 and 100 kD proteins...
Showing: 201 - 220 of 407 results