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We narrowed to 294 results for: expression vector

Showing: 41 - 60 of 294 results

  1. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ... A cis-splicing vector served as positive control (cis-ctrl). v1, vector 1; v2, vector 2; BD, binding ...SARS-CoV-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting microglia ... offers vectors for CROPseq-multi using selection by Puromycin, Zeocin®, or nuclear expression of BFP2...voltage sensors By Brian O’Neill New viral vector preps expressing the Voltron2 voltage indicator, created...administering these AAVs with a dilute Cre-expressing vector resulted in a sparsely-labeled field of view...high microglia-specific gene expression when incorporated in an AAV vector (Figure 5). Efficiency and specificity...of miR124T was inserted into the vector to silence transgene expression in neurons and other non-microglia...

  2. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...variety of pExpression vectors) plasmids. The type IIS restriction sites used in these vectors are of low...easier to produce the constructs you need. Expression vectors within this kit contain a variety of promoters... oocytes. The expression vectors additionally contain various combinations of protein tags (His, Avi, ... with the problem of creating vectors for predictable protein expression in bacteria, researchers at the...Process with pCri Cell-free protein expression kit Cell-free expression systems have been improved for both... the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of proteins in ...comparison of different cell-free expression systems; the Cell-free expression test kit represents a set of...

  3. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ...large packaging capacity and stable expression of lentiviral vectors make them an attractive option. However...using different types of AAV. Expression Another advantage of AAV vectors is how the cargo — in this case...multi-plasmid system, where you have one AAV vector dedicated to expressing Cas9, while the other houses the gRNA...However, safety requirements and expression limitations may be too constraining for some experiments, ... particularly in vivo ones. In those cases, AAV vector screens may be AAVantageous!       In vivo, in ...delivery method for in vivo CRISPR screens. AAV vectors can be made at high titers equal to those achieved...improved safety profile, customization, and enhanced expression in many mouse tissues. Safety AAV and lentivirus...

  4. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    Published
    May 28, 2020, 1:15 p.m.
    ...Lentiviral preps for expressing SARS-CoV-2 proteins ChR2 for targeted expression DREADDs GFP dependent...preps SARS-CoV-2 protein expression The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins ...2xStrep-IRES-Puro: Expression of the SARS-CoV-2 ORF3a Lentiviral preps for expressing the E protein (pLVX-EF1alpha-SARS-CoV...for targeted expression. Plasmid 75470 is now available in AAV1 for Flp-dependent expression of ChR2(H134R...GFP expression patterns, thus allowing you to further restrict gene activation to cells expressing both...pAAV-EF1a-Flp-DOG-NW Connie Cepko Viral vectors coming soon! These vectors listed below should be released at... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...

  5. 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...

  6. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    Published
    Nov. 27, 2019, 2:03 p.m.
    ...with AAV vectors are common approaches to control transgene expression by restricting expression to genetically...Addgene blog Download the viral vectors 101 eBook Read our viral vector related blog posts Learn more about... more. With the help of adeno-associated viral vectors (AAV), neuroscientists can combine classical genetic... how these changes affect the phenotype. Viral vectors and genetic approaches are continuously refined...designing experiments with AAV: 1. Getting your viral vector into your cell of interest: AAV tropism and route...Manipulate your cell of interest: Cell-type specific expression of molecular tools and sensors Combinations of...genetically defined cell types. Specific transgene expression can be also achieved in nontransgenic animals...

  7. 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...

  8. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    Published
    April 27, 2023, 1:15 p.m.
    ...your system. Chemogenetic vectors  Expression of retrograde serotype vectors is present at both the injection...-dependent vectors, this can cause challenges for the experimental design as these vectors can exhibit...phenomenon is to use a retrograde recombinase-expressing vector (e.g. Cre or FLP) to transduce the projection...tracing, but often at the cost of toxicity, low expression, or poor scalability.  Scientists at Janelia ...infects neurons, directs high-level transgene expression, and has retrograde functionality to monitor ...site while also activating or inhibiting gene expression, detecting neuronal signaling, or manipulating...AAV into Broca’s area of a mouse specifically expressing Cre in the hippocampus (Fig. 3). The Cre-dependent...

  9. 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...If one prefers expression in E. coli, the use of a thioredoxin A (TRX)-containing vector can improve disulfide... al. "The pCri system: a vector collection for recombinant protein expression and purification." PloS ...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...not fold well, you should screen a few different vectors to see which gives you the most functional protein...pCri also includes plasmids designed for protein expression in systems other than E. coli, specifically the...

  10. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...blog Viral Vectors 101: Viral Vector Elements Viral Vectors 101: Types of Viruses Viral Vectors 101: Viral...would not be great for a gene therapy vector! For viral vectors, only the protein for reverse transcriptase...components of the vector particle, some of the RT protein gets packaged inside of the vector particle, along...and this is where viral vectors and natural viruses diverge. For the viral vector, its job as a tool is ...transgenic genome will simply express from whatever promoters were included in the vector (or are nearby in the...instead get expression of the sensor, marker, or whatever other tool you chose to express. In the case...seem like an unlikely candidate to use as a viral vector, but retroviruses have been developed into some...

  11. 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 ...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...than viruses produced in cell lines that do not express these sugars. (Takeuchi et al., 1997). The alphaGal...

  12. 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 ...

  13. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...blog  Viral Vectors 101: Biosensors Viral Vectors 101: Viral Applications Viral Vectors 101: An Introduction... for neuron-specific expression (hSyn promoter) and pan-mammalian cell expression (CAG promoter). An improved...mammalian cell expression, and compatible with constitutive or Cre-dependent expression. These sensors...jGCaMP7 and jGCaMP8.   Viral vectors So, what do viral vectors have to do with all of these cool ...also already packaged into viral vectors and ready to go. Pro tip: Vectors with ready-to-use AAV preparations...can be more difficult due to variance in sensor expression if you aren’t working with a stably integrated...named CaMPARI2 is available for neuron-specific expression in both zebrafish and mammalian systems. The ...

  14. 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...

  15. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ... 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...

  16. 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...

  17. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    Published
    Sept. 17, 2019, 12:40 p.m.
    .... 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...

  18. 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... 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...

  19. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...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...

  20. 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...
Showing: 41 - 60 of 294 results