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 407 results for: expression vector

Showing: 61 - 80 of 407 results

  1. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ... non-recombinant vectors (re-ligated empty vectors, for instance) would still express ccdB and therefore... years ago and developed cloning vectors to harness it. These vectors, called pKIL18 and pKIL19, contained...coli that were transformed with the empty vectors expressed the ccdB gene and were therefore unable to...insert into the vector, the ccdB reading frame would be disrupted allowing cells expressing the recombinant...cells will not propagate while expressing the gene. Briefly, the vector “backbone” in this system contains...cells expressing it? The answer lies in special strains of E. coli that tolerate the expression of the...ccdB makes cloning easier by selecting against vectors that did not take up your insert. But exactly how...

  2. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ... ubiquitous expression of sgRNA(s) and mCherry (vector 1) with neuron-specific expression of dCas9-VPR... (i.e., how many sgRNA expression cassettes will fit). Cre-dependent expression of the SPH activator paired...system with ubiquitous expression of sgRNA (top) and neuron-specific expression of dCas9-VPR (bottom)....without inducing expression of the non-targeted variant. Moreover, isoform-specific expression of Bdnf resulted...differentially expressed genes between Bdnf I-overexpressing neurons and Bdnf IV-overexpressing neurons. Among...optimized vectors to highlight a key advantage of CRISPRa over traditional cDNA overexpression: isoform-specific... of precise genetic tools for modulating gene expression in the central nervous system (CNS). To address...

  3. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    .... Addgene has several recombinase-expressing ready-made AAV vectors which can be used in numerous floxed...researchers found that injecting lentiviral vectors expressing BDNF in the hippocampus causes a shift towards...studies using viral vectors in behavioral neuroscience. With the ability of viral vectors to fine tune gene...all of our viral vectors blog posts Find tips for getting started with viral vectors for behavioral neuroscience...IL-1 beta overexpression cassette. Expression of Cre in these neurons turns on overexpression of IL-1 beta...is activating, inhibiting, or modulating. Viral vectors, in contrast, are considerably more versatile and...behavioral tests, but now we’ll focus on how viral vectors have changed the scope and power of three commonly...

  4. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...transgene expression. Once packaged in AAV, this pre-recombined DNA could lead to transgene expression in recombinase-negative... their AAV vectors to find the optimal dose that allows for sufficient transgene expression but limits...blog Check out our Viral Vectors Featured Topic Page Subscribe to Viral Vectors Blog Posts Learn How We... of poor reagent quality. To make sure our AAV vectors are of the highest quality, we undertake a rigorous...To make sure that adeno-associated viral (AAV) vectors available at Addgene are suitable for in vivo studies...serotype. Additionally, we check viral transgene expression in transduced cells whenever possible. This post...should you care about viral quality control? AAV vectors are produced by scientists - not robots. Unfortunately...

  5. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...targeting vector (Fig. 1, Step 4), and co-injection of the targeting vector and a Cas9 expression plasmid...reaction to generate high-throughput targeting vectors. These vectors can be used to tag endogenous genes and...site destination vectors for inserting a tag at multiple target sites. SapTrap vectors can be easily modified...cells. To use CRISPaint, users need 3 vectors:  1) a gRNA vector that targets the gene of interest, 2)...then mixed and transfected along with a Cas9 expression vector. They can also be introduced directly into...cell line stably expressing Cas9. This PCR toolkit offers C- and N-terminal tagging vectors (eg GFP, Flag...PCR or cloning, as digestion of the destination vector with SapI yields 2 sites- the first site accepts...

  6. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...terminals with one vector. AAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun Luo’s lab expresses membrane tethered...element containing vectors developed in the Jordane Dimidschsteind lab provide expression selectively in ...interneurons. We now provide ready-to-use AAV vectors expressing GCaMP6f with this promoter. Image calcium...dendritic neurites from axonal neurites with AAV vectors expressing cytoplasmic targeted tdTomato and pre-synaptic...this method, the major capsid protein VP1 is co-expressed with a fusion protein containing VP2C and a protein...These plasmids with different promoters driving expression of Cas9 were used in this study.  The zCas9i ... a plant cloning kit that contains a series of vectors and DNA modules for assembly of CRISPR/Cas9 constructs...

  7. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...elements for vector propagation. This post, along with a future companion post on mammalian vectors, will catch...typically found as a single copy. Unlike the ARS vectors, CEN vectors are stable without integration. Yeast Episomal...of marker gene expression is to use a partially defective promoter to drive expression of the selection...plasmid maintenance within an E. coli cell, but vectors can be widely utilized across many different cell...core replication and resistance features of yeast vectors and explain how they differ from the bacterial ...previously described. Why do scientists use yeast vectors? Yeast are eukaryotes and thus contain complex ...facilitate simple gene replacement/mutation. Yeast expression plasmids used in the lab typically contain all...

  8. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...bacterial expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT...custom-cloned vectors in a Multisite Gateway reaction with a desired destination vector expressing a separate...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ... SFFV) or inducible (CMV-TO) cDNA expression, sgRNA (U6) expression, shRNA/shRNA-miR30 constitutive (H1...-miR30 (CMV-TO) expression, or a promoterless construct. Pre-constructed Entry vectors containing Cas9...a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed with the...enhancing target gene expression anywhere from 87x to 20,000x over endogenous expression (depending on the...

  9. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...to reduce basal expression levels; expression vector cannot have p15A origin of replication F- ompT lon...) pLysE has higher T7 lysozyme expression than pLysS; expression vector cannot have p15A origin of replication...Rhamnose-tunable T7 RNAP expression alleviates inclusion body formation. Expression vector cannot have p15A origin...for recombinant protein expression. Many challenges can arise when over-expressing a foreign protein in ...to avoid them. Why do I need an expression strain? Protein expression from high-copy number plasmids and...Addgene's curated list of Bacterial Expression Systems Table 1: E. coli expression strains  Note: All strains ...General protein expression BL21 (DE3) pLysS* Chloramphenicol (pLysS) pLysS expresses T7 lysozyme to ...

  10. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...protein expression constructs fused with epitopes or fluorophores, generate expression vectors with various...AAV transfer vector for in-vivo expression and plasmids optimized for dCas9-VPR expression in drosophila...parasites that constitutively express Cas9 with a library of sgRNA expression vectors (containing ten guides ... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ... to test multiple expression systems before they achieve the appropriate expression level for their gene...various promoters suitable for in vivo expression, and/or produce vectors to make mutant cell lines or transgenic... lab’s own pAc-sgRNA-Cas9 insect expression backbone, which expresses the gRNA from a Drosophila U6:2 ...

  11. Synthesized by Ginkgo Bioworks, Shared by Addgene: SARS-CoV-2 Plasmids for Many Expression Systems

    Type
    Blog Post
    ...cerevisiae expression vectors are available at Addgene. A set of mammalian expression vectors containing... be easily expressed in non-mammalian cells. Yeast expression plasmids The yeast expression constructs....  Bacterial expression plasmids For those constructs to be used in bacterial expression systems, Ginkgo...backbones which drive expression using either TEF1 or GAL1 promoters. These vectors are intended for the...tag, cleavage, codon usage, and expression system. Mammalian expression plasmids Finally a set of wild ... (spike protein) cloned into a CMV driven expression vector are also available. These contain no tags ...coding genes in E. coli and S. cerevisiae expression vectors in late March. The first set of 383 constructs...

  12. Using Addgene's New Viral Service for Your Research

    Type
    Blog Post
    ...Expanding the viral vector knowledge base: You can help! We’ve been expanding our viral vector educational resources...Beginners Lentiviral Vector Uses and Overview AAV: A Versatile Tool for Gene Expression in Mammals Additional...spending time and money producing virus from select vectors in the repository yourself, you can now order ready-to-use... production process while still ordering viral vectors. Virus production can be a frustrating process ...Lentivirus As some of the most popular lentiviral vectors in the repository are used to produce CRISPR tools... to knockout a gene, replace it, modulate its expression, or perform a genome-wide screen, we’ve got viruses...experiments, while rtTA can be used to activate expression of a gene under the control of the tet operator...

  13. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...recall, are the outer shell of an AAV vector, which determine the vector’s tropism, including its ability to...be paired with Cre-expressing transgenic mouse lines to restrict transgene expression to specific tissues...all take up precious cargo space in AAV vectors. Dual expression systems, which use two or more AAVs to...payloads. Since most systemic vectors are primarily derived from AAV9, the vectors have to be administered ...on the Addgene blog Viral Vectors 101: AAV Variables That Matter Viral Vectors 101: An Introduction to ...Selection of an Efficient AAV Vector for Robust CNS Transgene Expression. Molecular Therapy - Methods ...be suitable for your experiment. Like all viral vectors, the suitability of a systemic capsid for an individual...

  14. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ... of expression. For example, CAG is a strong promoter that drives very high, ubiquitous expression, while...outside of the ITRs does not get packaged in the AAV vector. The cloning capacity between the ITRs is ~4.4 ...of interest. Promoter The promoter drives the expression of the AAV transgene. On a plasmid map, promoters...type-specific, which can help you restrict the expression of your transgene to a specific species or location...to spatially and temporally control transgene expression. Lox sites are directional 34 bp sequences that...nuclear export of mRNA and helps increase the expression of your transgene. The WPRE is located at the...design a construct for the successful and specific expression of your gene of interest. References Haery ...

  15. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...that have excised the vector backbone. Vectors pTOX4, pTOX5, and pTOX6 express the YhaV, MqsR, and Tse2...excision of the vector backbone. To do this, rhamnose is added which induces expression of the toxin. Any...encode the amilCP chromoprotein. Vectors pTOX7, pTOX8, and pTOX9 express the YhaV, MqsR, and Tse2 toxins...gfasPurple) (Figure 2). Figure 2: Chromoprotein expression from bacteria on agar plate. Image: Liljeruhm...intensities, maturation times, and fitness cost of expression. They found that the cellular fitness cost varied...lost color overnight in liquid culture, though expressing the chromoproteins from the chromosome could ...allelic exchange Figure 3: pTOX plasmids expressing amilCP and tsPurple genes. Image: Lazarus et ...

  16. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ...Vital component for expression vectors: determines which cell types the gene is expressed in and amount of... referred to as “vectors” or “constructs.” To insert a gene of interest into a vector, scientists may ...plasmid with its elements described below.   Vector Element Description Origin of Replication (ORI...sites allowing for the easy insertion of DNA. In expression plasmids, the MCS is often downstream from a ...Regardless, once the cloning steps are complete, the vector containing the newly inserted gene is transformed...Generally, scientists use plasmids to manipulate gene expression in target cells. Characteristics such as flexibility...common plasmid types include cloning plasmids, expression plasmids, gene knock-down plasmids, reporter ...

  17. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors ... integrate into the host genome or temporarily express whatever it encodes. This ability to infect cells... Viruses are essentially DNA (or RNA) delivery vectors, so what makes them better than the alternatives...measures in a separate, upcoming post. If a viral vector seems like the right approach for your experiment...are generally used to achieve transient, high expression of the relevant cargo without the fear of random...Virus Pros Cons Retrovirus Stable Gene Expression Broad Tropism Only Infects Dividing Cells...Little Risk of Insertional Mutagenesis) High Gene Expression Infects Non-Dividing Cells Large Packaging Capacity...

  18. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...counterselection vector is the pDS132 sacB-based allelic exchange vector. This vector contains sacB, an...Tight control of toxin expression is absolutely necessary, because leaky expression of toxin could select...introduce a mutation in the chromosome is to disrupt expression of a gene with an antibiotic resistance cassette...in the chromosome, sometimes interfering with expression of surrounding genes. However, there are ways...selecting for an antibiotic resistance marker from the vector (abR, Figure 1). The resolution step (“double crossover...leads to the desired mutation and excision of the vector backbone. This step requires “counter-selection...colonies isolated are those where the allelic exchange vector has undergone single crossover and integrated into...

  19. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...About Viral Vector Elements Learn How to Do Genome-Wide Screens with CRISPR Lentiviral Vectors Additional...often employed to establish cell lines that stably express a gene of interest. While the general approach ...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting...compete with host cells for nutrients and can alter expression of receptors, ion channels, and growth factors...on a variety of factors such as the transgene expressed, the viral storage buffers used, and other experimental... by the envelop protein. Many times lentiviral vectors are engineered with non-native envelopes, a process...will be able to overcome differences in receptor expression. It's all in the mix Even before the lentiviral...

  20. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...engineered into viral vectors, so if you want to use them, you’ll need to use the vectors (or redesign the ...analyte), and accumulation of ATP activated GFP expression (receptor), then the transducer could be a microscope...retrovirus, can either permanently integrate and/or express long term (up to several years!). Biosensors tend...biosensor to specific tissues or cell (sub)types for expression and downstream monitoring. Nervous about making...Biosensors Viral Service: Biosensor AAV Viral Vectors 101 eBook Resources on the Addgene blog An Overview... as stable systems, rather than transiently overexpressing at the time of use. Another great viral perk...
Showing: 61 - 80 of 407 results