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We narrowed to 407 results for: expression vector

Showing: 21 - 40 of 407 results

  1. Multiplex Genome Editing with CRISPR-Cpf1

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    Blog Post
    ...than one vector to express the gRNAs and Cas9. Co-transfections can lead to variable expression levels ...larger expression vectors which are often more difficult to transfect or package into viral vectors. Cas9...Multiplexed Lentiviral Expression Cassettes lentiviral transduction One vector expresses Cas9, eGFP, and ...multiplexing vectors are larger because they require regulatory sequences to allow for expression of multiple...Lab Golden Gate Assembly transfection One vector expresses Cas9 and up to 7 gRNAs Each gRNA requires...Multiplexing crRNA expression with Cpf1  The key advantage of multiplexing crRNA expression with Cpf1 is that...vitro and when expressed in 293 cells. This allows for a single promoter to drive expression of a crRNA array...

  2. Plasmids 101: The protein expression toolbox

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    Blog Post
    ...permanently induce expression? Check out our Retro expression vector and Lenti expression vector. Turn it on...induce expression? Check out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off...ablate protein expression. If you need the ability to control when you turn off expression for good, the...knock out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for a little while Transient...inducible expression (more on this below) will both achieve temporary repression of protein expression. Addgene...appropriate promoter to express your cDNA, depending on the level of expression you would like. Check out...Addgene's Lentiviral dTAG cloning vector and SMASh degron tagging vector! Destabilizing domains Destabilizing...

  3. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

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    Blog Post
    ... gene is cloned into a viral shRNA expression vector and the vector particles are delivered into the brain...or lentiviral vectors in our previous blog posts.  Viral vector injection Viral vectors allow a tight ...using viral vectors By modulating neuronal gene expression and activity, you can use viral vectors to study...modification? and 2. Which viral vector should be used? Modifying gene expression Generally speaking, one may.... On the other hand, lentiviral vectors are superior packaging vectors for larger genes - they can hold...around 1012-1013 vg/ml (vector genomes per milliliter) and lentiviral vectors around 108 IU/ml (infectious...may be considered. Timing: High expression levels of the viral vector trangenes typically occur two weeks...

  4. Your Lentiviral Plasmid FAQs Answered

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    ... including cDNA expression, shRNA-mediated knockdown, Tet and Cre-regulated expression, CRISPR genome ...ingredients: the lentiviral packaging vector, a transfer vector including the gene of interest, and the... to drive expression of the gene of interest. Consequently, the levels of protein expression tends to ...3rd-generation transfer vectors apart? A3: Addgene defines 2nd- or 3rd-generation transfer vectors based on whether...between 2nd- and 3rd-generation vectors? What does this mean for transfer vectors? A4: The lentiviral packaging...generation of the transfer vector. Q5: What envelope glycoprotein will be expressed on virus particles? A5...cytotoxic precluding the long-term expression of lentiviral vectors in producer cell lines. Identifying...

  5. Plasmids 101: Mammalian Vectors

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    Blog Post
    ...using synthetic vectors and cultured mammalian cells. Of course, these mammalian vectors must be compatible...components expressed in trans within the cell for effective replication. Cell lines expressing the Epstein–Barr...straightforward and the resultant cells can either express the plasmid DNA transiently (similar to bacteria... easily monitor transfection efficiencies and expression levels in the cells. Ideally, the chosen reporter...reporter is unique to the cell, is expressed from the plasmid, and can be assayed conveniently. A direct... sufficient for the transfected plasmid to be expressed transiently. Since the DNA introduced in the transfection... into their genomes. Unlike plasmids used for expression in yeast and bacteria, plasmids used for stable...

  6. Plasmids 101: Shuttle Vectors

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    Blog Post
    ... as a shuttle vector? Are viral vectors shuttle vectors? The production of viral vectors typically involves...their vectors in fast-growing bacteria first. In fact, most yeast vectors and many mammalian vectors are...shuttle vectors are involved in the production of viral vectors, we consider the viral vectors themselves...between species. Figure 1: Shuttle vectors are vectors that can function in more than one host species...Not quite. Shuttle vectors have to address several big differences in the ways vectors are used across species...replication, selection, and expression. Replication It’s usually not enough to get your vector into your target ...needed to retain the vector as cells divide. In transient transfection, the vector remains episomal, meaning...

  7. 15 Years of Addgene: The Top 15 Plasmids

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    ...  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David Root lab is... 2nd or 3rd generation lentiviral vectors and an envelope expressing plasmid, such as our second most ...plasmid from the Zhang lab is a mammalian vector that expresses both SpCas9 and your gRNAs and was also ...(PX461) expresses both a Cas9 protein and a gRNA. But instead of Cas9, this plasmid expresses Cas9n (a...pSpCas9(BB)-2A-Puro (PX459) - This mammalian expression vector encodes both SpCas9 and your gRNA. It’s the...opening our UK office in 2014, to starting our viral vector service in 2016...we’re excited to help scientists...packaging plasmid from the Didier Trono lab that expresses the HIV gag, pol, rev, and tet genes. It can be...

  8. Viral Vectors 101: Viral Applications

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    ...interest, turning expression off permanently. Viral vectors can also be used to express proteins. Viral...Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors ...applications Gene expression control Researchers are always wanting to manipulate gene expression – turn it up...interest, and often a selectable marker. This vector can express your construct in target cells transiently... of these expression control mechanisms require a specific modification of the viral vector for your gene...We offer a variety of viral vectors, viral services and even a Viral Vectors eBook to get you started. ... terrains. They are, in fact, a choice delivery vector from basic science research all the way to clinical...

  9. Viral Vectors 101: Transductions

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    Blog Post
    ...Production at Addgene Viral Vectors 101: What is a Virus? Viral Vectors 101: Viral Vector Elements ...and viral particles are packaged, when the vector is expressed. Once the design and construction steps are...integrated into the host genome? Expressed episomally long-term? Expressed short-term? Answering these questions...nucleic acid cargo of your choice. First, a viral vector is selected to perform the transduction with: either...desired DNA to be integrated can be cloned into the vector that you have selected. The nucleic acid will be... it’s time to make virus! The constructed viral vector containing the cargo of interest is then typically...293 cells. When all of these viral plasmids are expressed within the production cell line, viral particles...

  10. Top Requested Lentivirus and AAV of 2016

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    ...blast LentiCas9-blast is a Cas9 expression vector developed by Feng Zhang’s lab at the Broad Institute... is a dual-vector system, meaning it is typically used in conjunction with a second vector that delivers...lentiGuide-Puro. The alternative single-vector systems typically express Cas9 and a gRNA from the same plasmid...constructs (i.e., using a dual-vector system as opposed to a single-vector system) has several benefits....stable Cas9-expressing cell line and then transduce this line with lentiviruses expressing various gRNAs...transductions with a single-vector system). Tips for using this plasmid Cas9 expression is typically cytotoxic...and transgene expression across different cell types (Aschauer et al., 2013). Expression of hM4D(Gi)-mCherry...

  11. Top Requested AAV of 2017: pmSyn1-EBFP-CRE

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    ... and other viral vector tools in the years to come. We’ll make many more viral vector tools and resources... A Versatile Tool for Gene Expression in Mammals Subscribe to Viral Vector Blog Posts Additional Resources... Service Page Browse Additional Viral Vectors Find Viral Vector Protocols ... retrograde (the most popular form), this viral vector is used to deliver DNA encoding Cre recombinase...synapsin promoter. It can be used to drive Cre expression specifically in neurons. AAV Delivered Cre - ...further control where your gene of interest is expressed.  A cell-type specific Cre can provide you with...Cre-dependent mCherry line with a separate line that expresses Cre in neurons, but both of these options would...

  12. An Introduction to Adenovirus

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    ...Viral Vectors 101: Viral Applications Adenoviral Vector Production and Troubleshooting Viral Vectors 101...fact, of how adenoviral vectors work. What are adenoviruses?   Adenoviral vectors are commonly derived from...majority of adenoviral vector systems used in research, they are only expressed in the producer cells ...cells that are infected by the vector, only the transgene will be expressed, since the early genes are absent...encapsidate new virions) are not expressed. Note that replication-competent AdV vectors also exist and are used...adenoviral vectors all have a genomic capacity that is much higher than other viral vectors (tens of kilobases...retroviruses). The tradeoff is that adenoviral vector expression is very short-lived. It is worth noting that...

  13. Lentiviral Vector Uses and Overview

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    ...Lentiviral vectors are one of the most popular and useful viral vectors in the lab. Advantages of lentivirus...dividing and non-dividing cells. Lentiviral vectors are the vector of choice for many CRISPR applications,...and future) applications of lentiviral vectors! Lentiviral vector history and components In the early ...viral vector systems based on retroviruses like Moloney murine leukemia virus (MMLV). The vectors could...transgene expression, but they could only infect actively dividing cells. Another type of vector, based ... but without sustained transgene expression. To design a viral vector system that could do both, Addgene...showed that expression persisted for at least thirty days. Although adeno-associated viral vectors can also...

  14. Viral Vectors 101: AAV Variables That Matter

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    ...know what expression pattern you need, and the factors in your system that can affect expression, before...early gene expression, but can become methylated over time, resulting in decreased expression (Damdindorj...tyrosine-hydroxylase (TH), can limit which cells express your gene. Off-target expression can still occur (Fig. 3) - for...transiently express TH - so you’ll need to check for this in your system and then test each vector before ...timing of expression in your experiment. Some inducible constructs can exhibit leaky expression in the absence...one that optimizes gene expression where you want it and minimizes expression where you don’t. The key...on the Addgene blog Viral Vectors 101: An Introduction to AAV Viral Vectors 101: The AAV Retrograde Serotype...

  15. Viral Vectors 101: Inverted Terminal Repeats

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    ...blog Viral Vectors 101: Viral Vector Elements Viral Vectors 101: Pseudotyping Viral Vectors 101: Systemic...transgene expression and transduction efficiency (Shitik et al. 2023). Transgene expression can be altered...ITRs can be used to drive gene expression at varying capacities. AAV vectors containing the AAV2 ITRs can...Inverted Terminal Repeat Sequence Role in Vector Transgene Expression. Hum Gene Ther, 31(3–4), 151–162. https...important biological tool that allow scientists to express a gene of interest in a cell or animal model. AAV...and are required for replication, packaging, and vector persistence. They work in conjunction with the ...retained with the gene in a recombinant AAV (rAAV) vector (Figure 1). Everything else outside the ITRs gets...

  16. Plasmids 101: Control Plasmids

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    ...Knockdown the expression of Gene X using shRNA expressed from a plasmid   Figure 1: Expression Level of Gene... the empty vector control shows any effect of the vector/backbone itself on gene expression in your target...as the expression level of Gene X in cells treated with Plasmid A is similar to the expression level of...empty control vector would be Plasmid A sans shRNA, or backbone Y alone. The empty vector control allows...decreased expression of Gene X. Ergo the positive control(s) should decrease the expression of Gene X....replicate controls.  Transfection controls: empty vector and internal control A transfection control measures...any effects of the transfection itself (i.e., the vector used in transfection, transfection reagent, or ...

  17. 22 Hot Plasmid Technologies from 2014

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    ... existing transposon and vector systems and creating an all-synthetic vector that included only the elements... into a destination vector in a predefined order. Finally, the destination vectors are used with Multisite... eight entry vectors can be used for each Multisite Gateway compatible destination vector for a maximum... to Addgene, including lentiviral and AAV expression vectors for certain variants. Klapoetke et al.,...step, compatible Level 0 vectors are directionally assembled into a Level 1 vector creating a single transcriptional...Cre-based Brainbow-expressing cells. Autobow plasmids are all-in-one versions which express self-excising ... May 5;10(6):540-7. pCoofy vectors for optimizing protein expression Under the direction of Sabine ...

  18. Viral Vectors 101: Chemogenetics

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    Blog Post
    ...targeting receptor expression as a ligand and receptor pair. Targeting Receptor Expression Using chemogenetics...therapies when expression must be restricted to specific circuits. Methods for targeting expression involve ...on the Addgene blog New to viral vectors? Browse our Viral Vectors 101 blog posts Read blog posts about...target effects. For example, if your receptors are expressed in the brain, it is necessary to use ligands capable...different responses depending on what cell type it is expressed in. The most common target for chemogenetic manipulation...modify other properties such as endogenous gene expression, which can lead to changes in normal cell physiology... H, Fishman GI, Conklin BR (1999) Conditional expression and signaling of a specifically designed Gi-coupled...

  19. AAVs CREATed for Gene Delivery to the CNS and PNS

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    ...labeling is to inject a mix of AAV vectors that constitutively express a red, green, or blue (RGB) fluorescent...blue are the predominate colors expressed (figure 4c).   AAV Vector  Percent of Neurons Transduced   ... color expression into a two part system (figure 4d):  Part 1: A cocktail of three AAV vectors expressing...using AAV as a tool for gene expression in mammals. Subscribe to Viral Vector Blog Posts Additional Resources...   Adeno-associated viral (AAV) vectors are the most frequently used gene-transfer tools in the study...are easy to manipulate, 2) they have long-term expression; and 3) they have limited toxicity. However, ... technique, a pool of mutant AAV capsid (cap) expressing plasmids is generated. Diversity is introduced...

  20. Negative Can Be Positive: Open AAV Data with Addgene

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    ...AAV vectors. I wanted to use an AAV vector to express ChR2(H134R) in A2 norepinephrine expressing neurons...serotypes containing an expression cassette with an ubiquitous promoter to express GFP in a group of young...to AAV vectors, many from people trying to answer the exact same question I was: What AAV vector is best...evaluate how small changes in vector administration can influence AAV expression properties.  By providing...Working with AAV vectors in the brain and peripheral nervous system presents a special challenge to scientists... brain region or nucleus. Just optimizing AAV expression can take months (or longer), yet the process ...question and used it to publish!  Working with AAV vectors in neuroscience Let’s consider my first challenge...
Showing: 21 - 40 of 407 results