Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 263 results for: Cre expression vector

Showing: 101 - 120 of 263 results

  1. 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 ...

  2. 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...reproducible. Prior to creating transgenic cultures or plants, our transient expression method enables us ...MoClo system you can easily and quickly create customized vectors. For more information on MoClo for plants...reporter gene so that the transformation and gene expression can be tracked. During a transient leaf transformation...References D'Aoust, Marc-André, et al. "Transient expression of antibodies in plants using syringe agroinfiltration...

  3. 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...proviral DNA synthesis. Increases transduction efficiency and transgene expression. Psi (Ψ) RNA target...element; sequence that stimulates the expression of transgenes via increased nuclear export. LTR Subcomponents...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 ...was originally published in 2018, in the Viral Vectors 101 eBook, 1st edition.  References and resources...

  4. The Importance of High Titer for AAV Transductions

    Type
    Blog Post
    Published
    March 1, 2018, 2:11 p.m.
    ...a roughly 6-fold increase in expression [3, Figure 2]. Note how expression increases exponentially with...observable expression. Doubling this titer to 1012 resulted in expression in roughly 200 cells. Increase the ... produce expression. The higher the titer, the better. AAV transport and transgene expression It’s clear...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...

  5. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    Published
    March 3, 2020, 2:15 p.m.
    ... of gene expression and reduces toxicity as compared to previous Cre/lox or drug-induced Cre systems (... Figure 4: The C-terminus of Cre (CreC) and the N-terminus of Cre (CreN) are fused to the truncated Phytochrome...As the name suggests, CreLite combines the Cre/lox system with optogenetics. Cre activity is controlled...determined protein expression levels from different promoters, and inhibited gene expression using CRISPRi.... dimerize, bringing each terminus of Cre together to restore Cre activity. As soon as red light or PCB...proteins separate, inactivating Cre. This is critical for reducing Cre toxicity in long-term experiments.... In this system, cells express various fluorescent proteins controlled by Cre recombination. Each cell...

  6. 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...Figure 1. The RepCap plasmid and its products. Created with BioRender.com   Replication (rep) gene...

  7. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...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...technology has been used to optimize protein secretion and expression in the biotech workhorse yeast — K. phaffii...GoldenBraid Modular Cloning System and Creation of a Toolkit for the Expression of Heterologous Proteins in Yeast... 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 ...

  8. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    Published
    Sept. 10, 2015, 2:30 p.m.
    ...with an appropriate mouse line expressing a particular cre, viral expression of your opsin is no longer required...steps in the coming weeks. Expression of the optogenetic constructs Expression of your desired optogenetic...promoter-directed expression alone is extremely limited. Therefore, highly expressing, generic neuronal...promoter of the cre recombinase while still taking advantage of the robust expression via the promoter...Finally, expression of the opsin in a mouse line will likely be less than that of viral expression but may...stimulation. Testing opsin expression  Regardless of the method of expression of your opsin gene, it is...extremely high expressing promoters). Conduct a time course experiment in which you monitor expression at various...

  9. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    Published
    May 2, 2024, 1:15 p.m.
    ...feature of this system is a lentiviral gRNA expression vector (pRDA_526) that (1) enables efficient knockout...murine tumor model in vivo. In this screen, we used the pRDA_526 vector to knock out 30 genes putatively ...the system and utility of the pRDA_526 vector. Moreover, the screen identified putative novel regulators... vivo screens in naïve T cells. More broadly, this system can be used to perform in vivo screens in other...power of in vivo screens. Figure 1: Using FITS to perform in vivo CRISPR screens to assess T cell...FITS In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However...However, in vitro screens cannot faithfully model the complex spatial, cellular, metabolic, and cytokine...

  10. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ... it easy to create a single gRNA-expressing plasmid, but it can also be adapted to express multiple gRNAs...one 30-minute reaction. The destination vector and entry vector(s) are placed in a single tube containing... 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...

  11. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...bacteria that have taken up this vector, but it won’t tell you whether the vector contains the gRNA insert or...on a plasmid, creates a functional 𝛽-galactosidase. Blue-white screens rely on vectors where the gene...deleted, it creates a non-functional 𝛽-galactosidase. Complementing this mutation by expressing this deleted...X-gal allows you to identify cells that took up the vector and distinguish between plasmids that contain the..., you will see two bands: one representing your vector backbone and the other representing the insert....doesn’t contain the insert, you will just see the vector backbone band. Or, you can choose an enzyme that...specific to your insert or by using primers that are vector specific but amplify a potential insert. If you...

  12. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    Published
    Dec. 7, 2015, 6 p.m.
    ...in CRISPR genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate inversions...you can use to express your protein of interest. 4. Plasmids 101: Blue-White Screening Getting a lot of...researchers with many different ways to control gene expression. Get the full scoop from Addgenie A. Max Juchheim... of empty vector contaminants in your cloning experiments and tired of having to PCR many colonies to ...CRISPR/Cas9 and AAV Adeno-associated viral (AAV) vectors are useful for in vivo gene delivery, but have ...Browse Our CRISPR Resources Learn More about Viral Vectors Browse the Repository  ...community members. In addition we’ve experienced incredible growth with our readership expanding from ~10,000...

  13. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...of Bacterial Expression Systems. Check out our companion post describing protein expression strains, where...Mutation in a chaperonin gene Increases the stability of certain expressed proteins endA, endA1 Endonuclease...mutations Blue/white screening of clones   lacIq lac repressor overproduced, expression from pLac repressed...lab strains designed for subcloning or protein expression. If you were to include customized strains, the...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)....

  14. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    .../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...Experimental output is measured and quantified. Created with BioRender.com.   Optogenetic tools Types...passing ions against the concentration gradient. Created with BioRender.com.   Opto-G-protein coupled...

  15. The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years

    Type
    Blog Post
    Published
    Jan. 8, 2014, 3:10 p.m.
    ...requests grow for lentiviral packaging and expression vectors, including pMD2.G and psPAX2. Actually, these... In 2006, we saw an increase in popularity of empty backbones for shRNA expression, such as pLKO.1 puro... Our first popular "kit," Andrew Fire's set of vectors for C. elegans research was our most requested ...puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with the pLKO plasmids ...empty backbones such as pBABE-puro for retroviral expression and pLVTHM, a lentiviral plasmid for mammalian...was Addgene's first distributed plasmid and was created in the lab of my thesis advisor, Dr. Bruce Spiegelman...requested. Ubiquitin - One particular gene we saw increased interest in from 2008 to 2009 was ubiquitin. The...

  16. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    Published
    April 23, 2020, 1:15 p.m.
    ...the percent of recombination present in Cre-dependent vectors and a script that can confirm the serotype... while we think of the AAV vector genomes as containing just the expression cassette that we want, it ... our viral vectors related blog posts Read our blog post on AAV titers Browse viral vectors protocols ...accommodate adeno-associated viral vector (AAV) samples and created a simplified process, termed viral...methods for AAV quality control Historically, AAV vector QC has been sparse including only titration of ... of quality control is sufficient but for viral vector core facilities producing a large number of viral...demonstrated that NGS is an effective tool to confirm AAV vector genome identity. These studies used SMRT sequencing...

  17. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 3:42 p.m.
    ...Viral Vectors 101: An Introduction to AAV Viral Vectors 101: AAV Variables that Matter Viral Vectors 101...choice of AAV vector for your experimental system. AAV1 AAV1 was the first viral vector approved for gene...: Diagram of AAV components. Created with BioRender.com. AAV vectors have both distinctive capsids...Widespread and Stable Pancreatic Gene Transfer by Adeno-Associated Virus Vectors via Different Routes....A viral vector that can target specific tissues, even when administered systemically, without causing...causing disease? Recombinant adeno-associated viral vectors, or rAAVs, can sound almost too good to be true...specific cells. This protein shell packages the vector’s genetic material, and variations between serotypes...

  18. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ...Addgene  Choosing an expression system Choosing an appropriate expression vector is the first step in ...2007) ‘Improved Gateway Binary Vectors: High-Performance Vectors for Creation of Fusion Constructs in Transgenic...from the Tsuyoshi Nakagawa lab are binary gateway vectors and can be used for promoter swapping and generating...for visual selection of transformants based on expression of RFP in the seed (Emami, Yee and Dinneny, 2013...many insertions can cause artifacts from over-expression and it is likely that such a construct will be... ‘A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology’, Frontiers in Plant...sweet potato contains Agrobacterium T-DNAs with expressed genes: An example of a naturally transgenic food...

  19. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ... terminator will suffice. Many commercial expression vectors use double terminators to reduce unwanted...Regulatory Element Enhances Expression of Transgenes  Delivered by Retroviral Vectors. Journal of Virology. ...polyadenlyation. Mammalian expression plasmids are primarily used to create mRNA and the commonly used...Internal Polyadenylation Signal Substantially Increases Expression Levels of Lentivirus-Delivered Transgenes...Plasmids designed to express genes in a given host cell type are generally broken down into two broad...on promoter strength as a determinant of gene expression levels, the terminator also plays an important...variability in RNA half-life, and ultimately gene-expression. Polyadenylation is the post-transcriptional ...

  20. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...Transgene Expression in Cultured Cells Using Unpurified Recombinant Adeno-Associated Viral Vectors [Preprint...minimally purified secreted rAAV for rapid and cost-effective manipulation of gene expression in the CNS. Molecular....  In this helper-free system, HEK293T cells, expressing Ad E1A and E1B, can be transfected with your ...you’ll need to determine the titer of your viral vector solution. A variety of titering methods have been...about common AAV applications by reading our Viral Vectors 101 series!  This post was originally written by...the Stability and Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy Methods, 28(1), Article 1. ...(2021). Inactivation of Adeno-Associated Viral Vectors by Oxidant-Based Disinfectants. Human Gene Therapy...
Showing: 101 - 120 of 263 results