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We narrowed to 407 results for: expression vector

Showing: 101 - 120 of 407 results

  1. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ...Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector production...acids. This conjunction of vectors and exosome was given the name vexosomes (vector-exosomes), exosome AAVs...transfecting the producer cell line with an expression vector for CD9 (a known marker molecule for microvesicles... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...production, assembled AAV vector particles accumulate inside the producer cells and purification protocols...purification steps. It’s also possible for AAV vectors to be spontaneously released from the producer ...Harvard Medical School took a closer look at the AAV vectors that are released from cells during production ...

  2. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    ...strategies. In such vector-based gene delivery systems, the stochastic expression of XFPs is possible ...reduced expression level of XFPs, most likely by suppressing transcription. It is critical to express a minimal...labeling, Brainbow used the Cre-loxP system to express one of the three fluorescent protein (XFP) genes... stochasticity will result in a combinatorial expression of these three genes with different copy numbers...This is because the stochastic and combinatorial expression of fluorescent proteins is only possible at low...Tetbow In modern neuroscience, plasmid or viral vector-mediated gene delivery is more common than conventional...Poisson distribution. To overcome the limited expression levels of XFPs in the original method, we used...

  3. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...Addgene blog Viral Vector Elements The Retroviral Life Cycle Popular Retroviral Vectors and Their Use in...method utilizes a packaging cell line that stably expresses Gag-Pol and/or Env, eliminating the need to deliver...  Frequently asked questions about γ-retroviral vectors How do engineered γ-retroviruses differ from lentiviruses...reduce promoter interference and/or drive high expression of a transgene from an internal promoter. An ...Increases transduction efficiency and transgene expression. Psi (Ψ) RNA target site for packaging ...regulatory element; sequence that stimulates the expression of transgenes via increased nuclear export. ...was originally published in 2018, in the Viral Vectors 101 eBook, 1st edition.  References and resources...

  4. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...observe expression from Agrobacterium, but not the plant. For example, we have seen strong expression of the...us. Use a well working vector system This allows you to quickly assemble vectors and test different ideas...MoClo system. The vectors mentioned in this blog post above are ready to use vectors, but with the MoClo... transgenic cultures or plants, our transient expression method enables us to quickly screen and investigate...reporter gene so that the transformation and gene expression can be tracked. During a transient leaf transformation...system you can easily and quickly create customized vectors. For more information on MoClo for plants, check...References D'Aoust, Marc-André, et al. "Transient expression of antibodies in plants using syringe agroinfiltration...

  5. Lentivirus Production

    Type
    Protocol
    ...Lentivirus Production You may also like... Viral Vector Guides Mol Bio Protocols Introduction This protocol...be used to produce lentivirus from a lentiviral vector transfected into 293T cells using a polyethylenimine...Figure 1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI...

  6. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    ...drives Cap expression. Often Rep proteins from the AAV2 serotype are used to package vector genomes flanked...AAV (rAAV) is made in the lab, rep and cap are expressed from a separate plasmid in trans, freeing up space...packaging: Rep78, Rep68, Rep52, and Rep40. Rep gene expression is often driven by the AAV p5 or p19 promoters...Strohl WR (2017) Adeno-Associated Virus (AAV) as a Vector for Gene Therapy. BioDrugs 31:317–334 . https:/...for Targeting and Improving the Efficacy of AAV Vectors. Molecular Therapy - Methods & Clinical Development...Adeno-Associated Virus Genome Interactions Important for Vector Production and Transduction. Human Gene Therapy...

  7. New and Upcoming Viral Vectors - December 2019

    Type
    Blog Post
    ...pAAV.hSynapsin.SF-iGluSnFR.A184S Viral vectors coming soon! These vectors should be packaged and available ... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV we have released in the...only interpret their results, but also confirm expression in target cells. While the various AAV serotypes... can be used to achieve different patterns of expression, researchers often need to determine these transduction...under control of the CAG promoter for ubiquitous expression. (For more details on this technology, see our...

  8. The Importance of High Titer for AAV Transductions

    Type
    Blog Post
    ... produce expression. The higher the titer, the better. AAV transport and transgene expression It’s clear...produce any observable expression. Doubling this titer to 1012 resulted in expression in roughly 200 cells... roughly 6-fold increase in expression [3, Figure 2]. Note how expression increases exponentially with...infect a cell to achieve visible expression of the protein. However, expression of receptors may not need to...concentrate this type of vector to such a high titer when looking for effective gene expression. How do you achieve...to the nucleus where their gene cargoes can be expressed. The bottom line is that a lot of AAV particles...transduction. In another example, neuronal gene expression by an anterograde AAV was shown to drop off drastically...

  9. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    ...operon repressor, thereby allowing lac expression). When expressed, the β-galactosidase enzyme can break... methods. Positive selection vectors encode a gene which, when expressed, is lethal to the cell. Cloning...screening overview blog post.) *Note: Two Popular Vectors for Blue White Screening Available at Addgene Are... that encodes for the enzyme β-galactosidase. Expression of the lac operon is induced by lactose, and ... plasmid, creating an α-complementation cloning vector. When a cloning reaction goes to plan and your ...enough time for any intact β-galactosidase to be expressed and process x-gal into blue pigment (16-20 hours...pUC18 and pUC19, and pBluescript are a few common vectors. Blue-white screening is just that – a screening...

  10. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...systems you can use to express the iGluSnFR3 variants: pRSET: Bacterial expression vector, use with IPTG induction... expression. CAG-FLEX: CAG promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin...neuronal expression. hSynap-FLEX: Synapsin-1 promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP...transmembrane domain in the mammalian expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol...FLP-dependent expression GFAP: Glial fibrillary acidic protein (promoter). Good for Glial expression (not neurons...SF-iGluSnFR. This improved variant has higher expression levels in bacteria and produces stronger fluorescent...dynamic range, photostability, quantum yield, and expression. Create yellow (v82) and lime green (v857) variants...

  11. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    ...restricts expression to GABAergic interneurons. These vectors can also be used as the control vectors for experiments...and control AAV These fluorescent protein expression vectors can be used to visualize cells and to quantify...pAAV-hSyn-mCherry at Addgene. Viral vectors coming soon! These vectors listed below should be released at... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV we have released so far...for order please email [email protected] with the vector name and serotype you’d like to be notified about...

  12. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ...luciferase, cre/lox, yeast expression, worm expression, insect expression, plant expression, mouse targeting, ...biosensor affinity for calcium.”  Primary Vector Type The vector type should describe how your plasmid ...to be used. Choose from: mammalian expression, bacterial expression, lentiviral, retroviral, AAV, RNAi...indicate type in “Vector Type Other” column. For example, you may enter “donor template vector”. Cloning...plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival strain. For plasmids ...

  13. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...conditional gene expression, multiplexed CRISPR, and chromosomal integration of larger expression cassettes ...chromosomal integration and gene expression, the system includes a toolkit to express recombinant proteins within...the Rapid Design and Production of Diverse Expression Vectors.” ACS Synthetic Biology, vol. 6, no. 7, American... tools can be used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even...from individual transcription units (TUs) used to express your favorite gene-of-interest, all the way to ...with E. coli and/or cell-free systems. Protein expression in yeast MoClo is also for eukaryotes! The technology...has been used to optimize protein secretion and expression in the biotech workhorse yeast — K. phaffii (...

  14. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    ...to test for cell type-specific expression of GFP. Half (113) expressed in retinal cells four weeks post-injection...More than 90% of expression occurred in one type of cell More than 50% of expression occurred in one type...of targeted expression. Surprisingly, <1% of synthetic promoters replicated the expression specificity...restrict expression to a single cell type based on the overlap in cell types both promoters expressed in. ...promoter to drive expression of Cre-mCherry and a second promoter to drive expression of a Cre-dependent... reporter. Expression of GFP will only result in cells where Cre-mCherry is also expressed. If Cre-mCherry...Cre-mCherry expresses in photoreceptors and horizontal cells, and Cre-dependent-GFP expresses in horizontal...

  15. New Viral Vectors - March 2024

    Type
    Blog Post
    ...as seven vectors newly available in the AAV PHP.eB serotype. We've also added two viral vectors that can...these new vectors, and many more, in the table below.    Table 1: New ready-to-use viral vectors in the ...St-Pierre New viral vector pAAV-hSyn-JEDI-2P-WPRE AAV9 Biosensor St-Pierre New viral vector pAAV-CAG-RdLight1... Tian New viral vector pAAV-hSynapsin1-RdLight1 AAV9 Biosensor Tian New viral vector pAAV_hSynapsin_psychLight2...viral vector pAAV-nEF-Con/Foff DREADD Gi-mCherry AAV8 Chemogenetics Deisseroth New viral vector pAAV.CMV.PI.EGFP.WPRE.bGH...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share...share the 21 additions we've made to our viral vectors repository this year (and it's only March!)  In our...

  16. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ...makes AAV vectors especially amenable to correcting disease-associated mutations. AAV vectors maintain...control of gene expression and the selection of specific promoters may also help AAV vectors evade host immunity... Check out our Viral Vectors Featured Topic Page Learn about CRISPR-AAV vectors Learn about Addgene AAV...thereby correct the mutations using retroviral vectors. However, many diseases require techniques that...target cells has been challenging. A variety of vector delivery technologies have been investigated including...usability in clinical settings. On the other hand, vectors derived from AAV have recently gained popularity...profiles as well as other benefits. What makes AAV vectors great gene delivery tools? There are a variety ...

  17. Rabies and Neuronal Tracing

    Type
    Blog Post
    ...Control Gene Expression with FLEx Vectors Resources on Addgene.org AAV Vector for TVA Expression AAV for ... TVA to Cre-expressing neurons in a transgenic mouse line. By doing this, RABV expressing the viral protein...to visualize how neurons are connected. Viral Vectors as Neuronal Tracers Due to their ability to efficiently...achieved by using EnvA-pseudotyped RABV, and only expressing the EnvA receptor (TVA) in user-defined cells...its surface will only infect Cre-positive, TVA-expressing cells (Osakada et al., 2013). Ultimately, this... genome, so viral progeny will not necessarily express EnvA unless it is supplied in trans. Monosynaptic...conjunction with restricted TVA and G protein expression is a powerful development in neural tracing. ...

  18. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...one 30-minute reaction. The destination vector and entry vector(s) are placed in a single tube containing... create a single gRNA-expressing plasmid, but it can also be adapted to express multiple gRNAs. Addgene... appear in the final construct. The destination vector contains sites with complementary overhangs that...TGGA and 3’ overhang TCCG can be ligated into a vector containing those overhangs. Entry DNA overhangs...enzyme and ligase. Although the original destination vector + insert may spontaneously religate, this transient...efficiently clone up to 7 gRNAs into one destination vector, making multiplexing easy.  Disadvantages of Golden... step. If your genes of interest or destination vector contain multiple internal restriction sites that...

  19. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    .../cells expressing the optogenetic tool causing a change in cellular activity or gene expression. Experimental...control everything from electrical activity to gene expression, and even CRISPR based gene editing. But, how...hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine excitation and inhibition by using both...confounding issues associated with heterologous expression of engineered receptors. OptoXRs are great for...candidates for your experiments.  Wavelengths Most vectors have an optimal wavelength to use but can be activated...

  20. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...of Bacterial Expression Systems. Check out our companion post describing protein expression strains, where...lab strains designed for subcloning or protein expression. If you were to include customized strains, the...chaperonin gene Increases the stability of certain expressed proteins endA, endA1 Endonuclease I (nonspecific... clones   lacIq lac repressor overproduced, expression from pLac repressed more   LacZ β-galactosidase...transformation efficiency deoR constitutive expression of genes for deoxyribose synthesis Allows uptake...   TOP10 derivative. For propagating plasmids expressing the ccdB gene (important in Gateway cloning)....Streptomycin  HB101 derivative. For propagating plasmids expressing the ccdB gene (important in Gateway cloning)....
Showing: 101 - 120 of 407 results