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We narrowed to 263 results for: Cre expression vector

Showing: 61 - 80 of 263 results

  1. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...Gateway pDEST, and pBAD vectors for example. Protein expression from each of these vectors is controlled by ...resulting in different levels of expression from each vector; lower expression may be required if your protein... and cell free expression systems. Overall the general strategy for protein expression consists of transfecting...which expression system to use depends on several factors: The protein you are trying to express How ...system. Mammalian expression systems Mammalian cells are an ideal system for the expression of mammalian proteins...for mammalian expression utilize viral promoters (SV40, CMV, and RSV) for robust expression post-transfection...insect expression systems; baculovirus infected and non-lytic insect cells. Baculovirus expression systems...

  2. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    Published
    April 7, 2022, 1:15 p.m.
    ... 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...systems you can use to express the iGluSnFR3 variants: pRSET: Bacterial expression vector, use with IPTG induction...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... eGFP for sfGFP to create SF-iGluSnFR. This improved variant has higher expression levels in bacteria ...range, photostability, quantum yield, and expression. Create yellow (v82) and lime green (v857) variants...

  3. Rabies and Neuronal Tracing

    Type
    Blog Post
    Published
    May 29, 2018, 1:51 p.m.
    ...Control Gene Expression with FLEx Vectors Resources on Addgene.org AAV Vector for TVA Expression AAV for ...example, a researcher can deliver Cre-dependent TVA to Cre-expressing neurons in a transgenic mouse line...RABV expressing the viral protein EnvA on its surface will only infect Cre-positive, TVA-expressing cells...Addgene Blog AAVs CREATed for Gene Delivery to the CNS and PNS Advanced Uses of Cre-lox and Flp-FRT - ...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... genome, so viral progeny will not necessarily express EnvA unless it is supplied in trans. Monosynaptic...

  4. Easing the Protein Purification Process with pCri

    Type
    Blog Post
    Published
    June 19, 2015, 3:08 p.m.
    ... information, you can then pick an expression vector and expression system. If you find that your protein... al. "The pCri system: a vector collection for recombinant protein expression and purification." PloS ...protein might not fold well, you should screen a few different vectors to see which gives you the most functional... the secretory pathway to improve folding and disulfide bond formation. If one prefers expression in E...special E. coli strains designed for protein expression. Cultures are then lysed and the protein of interest...stability and solubility. Determining the best way to express one’s protein of interest can save a lot of time...pCri also includes plasmids designed for protein expression in systems other than E. coli, specifically the...

  5. Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering

    Type
    Blog Post
    Published
    Nov. 12, 2014, 3:54 p.m.
    ...and efficiency for all-in-one CRISPR/Cas9 vectors expressing up to seven guide RNAs (gRNAs). Sakuma said...cloning method to ligate expression cassettes of the gRNAs into a single vector in tandem. They went on...A newly established all-in-one vector construction system for CRISPR/Cas9-mediated multiplex genome engineering...systematically established way of making an all-in-one vector for multiplex genome engineering.” Now, they’...Schematic overview of the all-in-one CRISPR/Cas9 vector construction system for multiplex genome engineering...their CRISPR/Cas9-nuclease and CRISPR/Cas9-nickase vectors work successfully to target multiple sites in cultured...it has not yet been demonstrated, they say the vectors are also likely to work in animal embryos based...

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

    Type
    Blog Post
    Published
    July 28, 2017, 1:30 p.m.
    ...Learn about Using Cre in Drosophila Read Other Genome Engineering Posts Use FLEx Vectors for Site-Specific... 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...

  7. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...lentiviral vector will infect, pseudotyping can help. In pseudotyping, you produce viral vectors using viral...Sandoval S (2017) Pseudotyped Lentiviral Vectors: One Vector, Many Guises. Human Gene Therapy Methods ... Lentiviral vectors pseudotyped with a modified RD114 envelope glycoprotein show increased stability in...To deliver genes using lentiviral vectors, you need an envelope protein on the virus’s surface and a ...virus. How to pseudotype To pseudotype your viral vector, you use the same protocol for generating lentivirus...Sometimes this is due to the cells that the viral vectors were produced in. For example, production of vesicular...virus, HIV-2, and human foamy virus in cell lines expressing galactosyl(alpha1-3)galactosyl (alphaGal) sugars...

  8. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...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...

  9. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    .... At the time, adenoviral vectors were the most commonly used viral vectors for gene delivery, and they...transiently. However new viral vectors capable of providing long-term gene expression in retinal cells - AAV ... better vector for ocular gene therapy: Adeno-Associated Virus (AAV) While other viral vectors such as...immunogenicity and its transient expression. LV results in long term expression in dividing and non-dividing...technician in a lab using adeno-associated viral vectors (AAVs) to treat an inherited retinal degenerative...tolerate the presence of antigens (such as viral vectors) without triggering a destructive inflammatory ...transduce retinal cells, AAV quickly became the vector of choice for ocular gene therapy. AdV and LV ...

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

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...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...producing multiple vectors in parallel because the risk of accidental errors increases. In addition to human... 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...

  11. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...bacterial expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT...luciferase) to create a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed... to create the pCell-Free vectors, Gateway-compatible backbones which enable cell-free expression of proteins...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...reaction with a desired destination vector expressing a separate reporter gene (EGFP, iRFP, IFP1.4, puromycin...

  12. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...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 ...easily create protein expression constructs fused with epitopes or fluorophores, generate expression vectors...large-scale expression screen that can be used to generate His-tagged human kinase constructs that express well... 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...

  13. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    Published
    Jan. 11, 2017, 8:47 p.m.
    ...This is a versatile mammalian expression vector that you can use to express both SpCas9 and your gRNA in...this is a versatile mammalian expression vector that you can use to express both SpCas9 and your gRNA of...the top requested plasmid from 2016 is an expression vector for CRISPR nuclease, SpCas9 from the Zhang...mammalian cell types. Purpose  SpCas9 This plasmid expresses Streptococcus pyogenes (Sp) Cas9 from a Cbh promoter...resistance gene as separate peptides. PX459 This is the vector backbone. This particular backbone can be used ...N-terminus. This tag can be used to test for SpCas9 expression post transfection by western blot. Overall, this...mammalian cell lines. Alternative forms of this vector contain GFP-tagged and un-tagged versions of SpCas9...

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

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ... 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...

  15. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    Published
    Feb. 25, 2014, 7:11 p.m.
    ...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...to be created in advance of the experiment. The marker provided by the plasmid may be expressed at higher...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 ...

  16. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...multicolor labeling, Brainbow used the Cre-loxP system to express one of the three fluorescent protein ...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... 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. Synthesized by Ginkgo Bioworks, Shared by Addgene: SARS-CoV-2 Plasmids for Many Expression Systems

    Type
    Blog Post
    Published
    May 19, 2020, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...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 ...

  19. New and Upcoming Viral Vectors - December 2019

    Type
    Blog Post
    Published
    Dec. 19, 2019, 2:40 p.m.
    ...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...pAAV-hSyn Con/Fon hChR2(H134R)-EYFP that is both Cre and Flp dependent. Plasmid Serotype Name 124603...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...

  20. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...Yeast Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read...activation. Vary the expression levels of bait and prey proteins - overexpression can lead to false interactions...interactions. Lowered expression levels will increase the stringency of your binding. And most importantly...ends’ of your protein are screened for binding. Using a variety of expression levels and systems can alleviate...produce a more vigourous screen is to use a number of different bait and prey vectors. This has been shown...During this period, proteins were successfully expressed as fusions that retain their individual activities...see Figure 2a) (4). These fusions are then co-expressed in a yeast strain that also contains a reporter...
Showing: 61 - 80 of 263 results