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We narrowed to 294 results for: expression vector

Showing: 81 - 100 of 294 results

  1. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    Published
    Oct. 17, 2019, 12:28 p.m.
    ... optogenetic FLEx vectors in combination with transgenic animals or rAAVs expressing Cre under a specific...not naturally light-sensitive, so the selective expression of opsin genes in targeted cells makes it possible...How to get an opsin into the brain In order to express opsins within the brain, researchers inject genetically...relatively stable extra-chromosomally long-term expression and low ability to induce innate immune responses... promoters; however, those usually drive weak expression of downstream genes. Strong cell-specific promoters...Vandenberghe, 2019). How to achieve strong opsin expression in specific neuronal cell types then? This is... precise temporal and spatial control of gene expression in vivo (Schnütgen et al., 2003). This is achieved...

  2. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...Cas9. Adenoviral CRISPR vectors (left) or CRISPR vectors and donor DNA vectors (right) are transduced ...need to use a two viral vectors system: one vector for Cas9/sgRNA and one vector for your specific DNA ...gRNA and Cas9 using adenoviral vectors As compared to other viral vectors, AdVs can provide a large transgene...than AAV vectors. This capacity is large enough to carry the Cas9 gene and a gRNA expression cassette ...adenoviral vectors Since then, other groups have successfully developed and used AdV vectors to edit genes...updated Dec 10, 2020 by Benoit Giquel.   Adenoviral vectors (AdVs) have been used for many years for in vivo...of oncogenicity or genotoxicity seen with other vectors (Lukashev et al., 2016). The protein capsid encapsulating...

  3. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core. Transfections...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....Transducibility Some viral vectors are tested in vitro and in vivo for gene expression and/or function. These...Empty vector particles can be identified after negative staining and appear darker than full vector particles...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...to determine which facility produced your viral vector lot. At Addgene, AAV particles are titered by droplet...method.  Titering by the University of Pennsylvania Vector Core is (as of April 2016) also performed by ddPCR...

  4. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...target cells that are positive for vector encoded transgene expression following transduction with serial...target cell line with your virus and assaying for expression of a gene carried on the transfer plasmid or ...primers that target the common features of viral vector backbones; once validated these universal primers...higher than for functional assays depending on the vector backbone. When using qPCR based methods for viral...plasmid which can over-inflate results. A packing vector deficient in reverse transcriptase will produce... prep. This method is particularly useful when vectors carrying fluorescent markers are used as it eliminates...addition, promoter choice can also impact transgene expression. The most accurate method of lentiviral titration...

  5. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...: A knockout targeting vector designed to insert a resistance gene. The vector contains a neomycin resistance... Resources on Addgene.org Find Mammalian Expression Vectors Find Cre-Lox Plasmids Learn More about CRISPR... homologous sequence using a specially designed vector that contains sequence homology to the locus of... of the targeted gene have been cloned into the vector on either side of an antibiotic resistance gene...Figure 2). Figure 2: A knockout targeting vector designed to insert and later remove a resistance... the sequence to be inserted is cloned into the vector between the homologous sequences together with ...disrupted (knocked out) but the inserted GFP is expressed (knocked in). As seen in the above example, you...

  6. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...to create recombinant vectors that can replicate in different bacteria and express proteins within different... S. (2011). Broad-host-range plasmid vectors for gene expression in bacteria. Methods in Molecular Biology...Gill, R. T. (2010). Broad-host-range vectors for protein expression across gram negative hosts. Biotechnology...Cloning Vectors for Gram-Negative Bacteria. In R. L. Rodriguez & D. T. Denhardt (Eds.), Vectors (pp. 287... inform us on how to create better recombinant vectors. In this blog post, we’ll define plasmid host range...features to create broad host range recombinant vectors.  A plasmid’s host range is determined by the ...groups (Jain and Srivastava 2013).  Recombinant vectors that are made from broad host range plasmids are...

  7. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...Adenoviral vectors (AdV) are attractive vectors for research applications and gene therapy: they can ...Three generations of adenoviral vectors: the first generation vectors are deficient in the E1 and E3 regions... second generation vectors are deficient in E1, E2, E3, and E4. Gutless Ad vectors (helper-dependent (...creating adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the transgene...death of transduced cells and loss of transgene expression (Interestingly, the strong immunogenicity of ...Adenovirus type 5. Three generations of adenoviral vectors The adenovirus virion is a non-enveloped particle...capacity is limited to 8.2 kb, and in vivo transgene expression ceases relatively quickly due to immune response...

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

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...Description Expression Additional considerations CMV General expression mRNA Strong mammalian expression promoter...lower expression than U6. May have better expression in neuronal cells. U6 small RNA expression shRNA...some cell types. EF1a General expression mRNA Strong mammalian expression from human elongation factor ...give consistent expression regardless of cell type or physiology. SV40 General expression mRNA Mammalian...  Commonly used in expression systems for Drosophila. Polyhedrin General expression mRNA  Strong insect...Constitutive  Commonly used in expression systems for insect cells. CaMKIIa Gene expression for optogenetics mRNA...neuronal/CNS expression. Modulated by calcium and calmodulin. GAL1, 10 General expression mRNA Yeast ...

  9. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...System Application Mammalian expression vector Promoter driving Cas9 expression can be constitutive or inducible...included with each expression system as well as suitable applications. Expression System Components ... in a single lentiviral transfer vector or separate transfer vectors. May contain reporter gene (e.g. ... AAV transfer vector and used to generate AAV particles. Transient or stable expression of SaCas9 and/...or electroporation. Transient expression of CRISPR components, expression decreases as RNA is degraded ...using cationic lipids. Transient expression of CRISPR components, expression decreases as gRNA and Cas9 protein... provide an overview of some CRISPR mammalian expression systems, the typical applications for each, and...

  10. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    Published
    June 6, 2023, 1:15 p.m.
    ...Viral Vectors 101: Viral Vector Elements Viral Vectors 101: An Introduction to AAV Viral Vectors 101:...integrate into proto-oncogenes and disrupt their expression (Robinson, 1982). To add insult to injury, retroviruses...cells and doesn’t require genomic integration to express its cargo, in fact its viral life cycle does not...not include genomic integration. While the expression of adenovirus cargo is high, it’s generally not ... effective option if you need high, transient expression of a genetic cargo or have a very large cargo... Galla M, Baum C, Schambach A. Gammaretroviral vectors: biology, technology and application. Viruses. ...

  11. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    Published
    March 19, 2019, 1:31 p.m.
    ... cloned into an entry vector. The 2in1 cloning system contains two entry vectors (pUC57-L1L4 and pUC57... of protein coexpression. By introducing a single plasmid containing both GOIs, expression levels are ...-protein interaction studies easier with a 2in1 vector cloning system Article contributed by Michelle ... Recently, the Grefen lab introduced the “2in1” vector cloning system - a Gateway compatible approach ... genes-of-interest (GOI) into two independent expression cassettes on a single plasmid (Grefen and Blatt...digest cloning to introduce the GOIs into the 2in1 vector instead of the GatewayTM cloning BP reaction, where...Agrobacterium-mediated DNA transfer, each GOI is usually expressed from a different plasmid, which may lead to unequal...

  12. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    Published
    July 28, 2017, 1:30 p.m.
    ... plasmids for overexpression and knockdown. Addgene also has a number of insect vectors to consider. An... your gene of interest in a plasmid and get the vector incorporated into the germline. To begin this process... browse Addgene’s Collection under the insect expression system or by filtering by fly as species of gene...Find Gal4 and UAS plasmids at Addgene Choosing a vector to generate a transgenic fly There are a few important...important points to consider when choosing the vector you’d like to use to generate your new fly line:... gene to be incorporated into the genome? Some vectors incorporate into the genome randomly, while others...insertion has the advantage that you can get varying expression levels of your transgene in different fly lines...

  13. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...interference (RNAi) to knockdown expression of specific gene expression. The 49 deposited plasmids enable... all have a PR-T4 vector backbone, which is a modified version of L440. This vector is versatile in that...of interest into the empty PR-T4P vector backbone. The empty vector PLL4, which is similar to PR-T4P, ...Ingestion of bacterially expressed double-stranded RNA inhibits gene expression in planarians." Proceedings...Scientists routinely use techniques to alter gene expression or to label specific cells, but there are too... in Situ Hybridization (WISH) for visualizing expression of genes. Double-stranded RNA (dsRNA)-mediated...enable students to study gene expression and functions of key regeneration-related genes in four inexpensive...

  14. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    Published
    March 27, 2018, 1:32 p.m.
    ...selectable markers that are only expressed when the targeting vector is inserted downstream of a functional...them sequentially into the chosen acceptor vector. The vectors are then packaged into AAV virions via transient...Check out our Viral Service Find AAV Vectors Browse all Viral Vectors ...fantastic gene delivery vehicles for episomal gene expression and are particularly useful for gene delivery...well understood, researchers have been using AAV vectors for gene editing in mammalian cells since 1998....when delivered via AAV infection, gene targeting vectors with relatively short (1 kb) homology arms underwent...orders of magnitude higher than identical targeting vectors delivered through transfection of naked DNA. Other...

  15. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    Published
    March 14, 2019, 1:54 p.m.
    ...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 ...

  16. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...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...

  17. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    Published
    Aug. 6, 2024, 1:15 p.m.
    ...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...

  18. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    Published
    July 25, 2019, 12:03 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Dec. 3, 2020, 2:15 p.m.
    ...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...

  20. The Importance of High Titer for AAV Transductions

    Type
    Blog Post
    Published
    March 1, 2018, 2:11 p.m.
    ... 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...
Showing: 81 - 100 of 294 results