Skip to main content
Addgene

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

In Category

We narrowed to 409 results for: expression vector

Showing: 141 - 160 of 409 results

  1. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...to be expressed. These permit exquisite control of gene expression and tracing of gene expression across... basal expression. Co-expression of repressor QS along with QF leads to intermediate expression from QUAS...transgene permits directed expression of that gene in cells that also express GAL4. Usage of this system...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...basal state, and co-expression of the inducer QF can increase QUAS driven expression by ~3,300-fold in ... Shen. “Controlling gene expression with the Q repressible binary expression system in Caenorhabditis ...gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant...

  2. Hot Plasmids February 2024

    Type
    Blog Post
    ...escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in human cell lines...structured illumination microscopy in HeLa cells expressing COX8a-mStayGold. Scale bars 10 μm (main), 1 μm...developed an engineered adeno-associated virus (AAV) vector named AAV.CAP-Mac for non-invasive systemic gene...bias in mature marmosets. CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured...cultured human neurons, with broader neuronal expression than AAV9. This new capsid will be a valuable ...N.C., Goeden, N. et al. Adeno-associated viral vectors for functional intravenous gene transfer throughout...region VIII of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial cell-specific tropism, with...

  3. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    ...plasmids are formed. Viral vectors are prone to recombination In lentiviral vectors, recombination often occurs...Improved recombinational stability of lentiviral expression vectors using reduced-genomeEscherichia coli. BioTechniques...problems that I help troubleshoot involves viral vectors and plasmids with repeating sequences. Repeating.... Over time, bacteria containing the recombined vector backbone will also outgrow bacteria that contain...into two smaller plasmids. The ~3.7 kb recombined vector backbone contains the antibiotic resistance gene...samples? Plasmid recombination can affect viral vectors (LTRs and ITRs), plasmids with multiple polyA sites...recommendations from the depositing lab. If your vector is large (>10 kb) or has a low copy number, try...

  4. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...the vector backbone you cloned your insert into and fill out the appropriate boxes with the vector backbone...backbone manufacturer, vector backbone size, total size of your vector + insert, any modifications you ...insert in E. coli, then this is a Bacterial Expression vector. If your plasmid can be classified in multiple...lentiviral vector to be used in mammalian cells, check Lentiviral and Mammalian Expression). This information... location, while shRNAs are used to knockdown expression of mRNAs with complementarity to the shRNA sequence...provide any full or partial plasmid sequences and any vector maps. The more sequence data available, the better... was a part of the insert (when cloned into the vector backbone). If your plasmid has more than one insert...

  5. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    ... insert and vector are first PCR amplified. The forward primer used to amplify the vector is located in...your vector and insert. Once you’ve confirmed that the PCR was successful, mix together the vector and ... PCR with vector specific primers to confirm that the products are the correct size (vector + insert)....specific sequences for base pair overhangs, insert and vector purification, and, most importantly, excessive ..., it does not require purification of insert or vector products, nor does it require the use of specific...process in brief In FastCloning, you PCR both the vector and the insert in separate reactions and then combine...bases that are complementary to the overlapping vector ends. See an overview of the PCR process and the...

  6. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ...available in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). Fabritius...APOBEC. To achieve this they created a lentiviral vector system that monitors both APOBEC- and Cas9-mediated...ACE) in real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry...both halves of the modified EB1 protein are overexpressed in cells, microtubule growth can be switched...

  7. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...cloned into a vector containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector...repair, making it easier to construct targeting vectors. Addgene depositor Takashi Yamamoto’s lab has harnessed...just upstream of a stop codon. Briefly, the PITCh vector should be constructed with 5’ and 3’ microhomology...PITCh Step 1: Generate microhomologies in the PITCh vector ~20 bp 5’ and 3’ microhomologies are added to ..., and this construct is inserted into the PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an... vector carrying Cas9 and the PITCh- and locus-specific gRNAs Step 4: Select for puromycin resistant cells...plasmids available from Addgene are perfect for expressing GFP from a given promoter, and the technique ...

  8. What the HEK?

    Type
    Blog Post
    ...derivatives of HEK-293, the most commonly used line expresses SV40 large T antigen and is thus dubbed HEK-293T...packaging cells’ for viral production because they express the SV40 large T-antigen. This allows them to replicate...receptors and ion channels are often exogenously expressed and studied in HEKs due to the cell line’s ability...produce these proteins and its relatively low expression of native channels.  No matter what your research...factors that could skew research results (gene expression, growth rates, etc.). Thus, it’s important to...Platform to Produce Recombinant Proteins and Viral Vectors. Front Bioeng Biotechnol. (2021) (9):796991. DOI...10 Tips for Mammalian Cell Culture Adenoviral Vector Production and Troubleshooting Additional resources...

  9. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...because they can be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs... plant, and insect cells, they are most often expressed from a plasmid in bacteria (Ahmed et al., 2012...2012). One challenge of bacterial expression is the proper formation of the disulfide bond between the VL...requires an oxidizing environment to form. When expressed in the non-oxidizing cytoplasm of bacteria, scFvs...more of these approaches are often used for scFv expression in bacteria:  targeting to the oxidizing periplasm...to allow for proper disulfide bond formation.  expression in strains of bacteria that are redox mutants...mutants so they have a more oxidizing cytoplasm. expression in the presence of molecular chaperones that help...

  10. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...recognized by the Sep-tRNA. 3. An expression plasmid encoding the ORF to be expressed Finally, the researcher needs...ORF of interest is cloned into one of these expression vectors, the codon for the amino acid to be replaced...non-phosphorylated serine at the TAG codon of the same expression vector used for phosphoserine incorporation (6). ...bacterial host strain for protein production, and an expression plasmid encoding the open reading frame (ORF)...encoding their open reading frame of interest. Two expression systems have been verified for use by the Rinehart...TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing GFP that can ...using standard protein purification strategies; expression should be optimized for parameters such as induction...

  11. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    ...guide to viral vectors Find all of our viral vector blog posts Download our viral vectors 101 eBook Additional...pressure on the production of retroviral vectors: Enhancement in vector stability." Biotechnology and bioengineering...been actively investigating ways to improve viral vector production. Whether redesigning viral packaging...membrane When packaging adeno-associated viral vectors (AAV) not all serotypes are equal; while some serotypes... of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...we believe that this approach can be adapted by vector cores and laboratories alike as an easy and inexpensive...Efficient serotype-dependent release of functional vector into the culture medium during adeno-associated...

  12. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    ...codon optimized specifically for expression in yeast. These tagging vectors are based on previously described...interest. These fluorescently tagged genes are expressed under native conditions and allow scientists to...described pFA6a-link vectors and include a Kan, SpHIS5, or CaURA3 selection marker. Lee et al.(1) assessed...from these yeast-optimized fluorophore tagging vectors for your single or multi-color imaging experiments...artifacts sometimes associated with plasmid-based overexpression. Tim Formosa, at the University of Utah, has..., including full and partial gene deletion, overexpression (by promoter substitution), and tagging at ...

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

    Type
    Blog Post
    ... 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 ... platform to accommodate adeno-associated viral vector (AAV) samples and created a simplified process,...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... simplified approach, we sequenced over 100 AAV vector preparations and showed that VGS can quickly and...

  14. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    ...easy process. The cloning capacity of lentiviral vectors (~8.5 kb of insert) and the reduced titer associated...providing lentiviral preps of select lentiviral vectors from the repository for you. We perform test infections...infections with all of our viral preps and host our expression data (western blots or fluorescent microscopy...nickase for genome editing. You can also enhance expression of your gene of interest with a variety of Cas9...therefore, must be used to infect a cell line already expressing Cas9. As John Doench (Broad Institute and member...Page Check Out Our CRISPR Guide Pages Find Viral Vectors ...

  15. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...modular vector assembly process designed for a quick, plug-and-play approach to cloning CRISPR vectors (McGee...for each module, generate the vector file, and quickly clone their vector of interest. And it truly is ...destination vectors for various delivery methods, along with compatible inserts. All destination vectors contain...heavy lifting out of CRISPR vector design, making it easier to get the vectors that are just right for your...Fragmid destination vectors currently available. Table 1: Fragmid destination vectors Organism Delivery...designing a CRISPR vector the Fragmid way!    Figure 1: Selecting your vector type on the Fragmid...plasmid and generating the vector file on the Fragmid website.   If the vector is to your liking, your...

  16. Top Requested Plasmid of 2017 - pMD2.G

    Type
    Blog Post
    ...2016 were CRISPR plasmids), but so are lentiviral vectors, many of which are used to deliver Cas9 and other....G from Didier Trono’s lab! Lentivirus envelope vector pMD2.G This plasmid has been requested nearly 6,000... 6,000 times (over 800 orders in 2017) and expresses the VSV-G envelope protein under the control of the... our Viral Production Process Find other Viral Vectors ...

  17. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

    Type
    Blog Post
    ...pools are available in two vector formats: all-in-one (lentiCRISPRv2, expressing both Cas9 and an individual...individual gRNA from this single vector) and guide-only (lentiGuide expressing only an gRNA, for use with cells...plasmids will also be useful for imaging or gene expression assays necessitating the screening of individual...

  18. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    Blog Post
    ... injection of an AAV expressing a ChRger. In both experiments, neurons expressing ChRgers produced stronger... what was seen with ChRger2 expressing mice, while ChR2(H134R) expressing mice did not have this self-...Addgene.org Browse all viral vectors available at Addgene Visit our viral vectors guide Find our chemogenetics...to fire when light is shined on them, but the expression of light-gated ion channels such as channelrhodopsins...When light shines on excitatory channelrhodopsin-expressing neurons, the channelrhodopsins respond by opening...sequence and structures that determine membrane expression and plasma membrane localization of transmembrane...and ChRger3, were then characterized further by expression in cultured neurons and mouse brain slices collected...

  19. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ...multiplexed gRNA expression (Wong et al., 2016), in addition to the combinatorial expression of other genetic...CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12 ...below and pool-clone them into the pAWp28 storage vector (Figure 1): Forward oligo: 5’- CACCGNNNNNNNNNNNNNNNNNNNGTTTGGGTCTTCGAGAAGACCTATTCXXXXXXXXC...barcode, respectively. Once cloned into the storage vector, the pooled, barcoded gRNA library is now ready...pAWp12 lentiviral vector should retain the diversity in your gRNA library, but this should be verified by NGS...

  20. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...Fluorescent Protein Plasmids Find Mammalian Expression Vectors ...that overexpress genes of interest; more recently we have also been using Addgene CRISPR vectors to generate... enzyme assays to select both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected...and PCR, depending on the level of endogenous expression. We highly recommend the use of an electronic...ampules each of 4 clones with different levels of expression of our gene of interest. Advantages of the agarose...isolate individual cell clones with different expression characteristics from a population of transfected...Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org Check Out Our Protocols...
Showing: 141 - 160 of 409 results