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We narrowed to 294 results for: expression vector

Showing: 1 - 20 of 294 results

  1. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    Published
    July 25, 2023, 1 p.m.
    ...of viral vectors here! Need Virus? Check out Addgene's Viral Service! Types of viral vectors Of the many...types, but most vectors are based on adenovirus 5 (Ad5).     Fig. 3 - Viral vector components of...of adenovirus Building viral vectors Addgene has many viral vectors and helper plasmids for all the...adeno-associated virus (AAV). The components of these vectors are also split across several plasmids (often three...called helper plasmids. Below, we’ll breakdown the vector components first by virus and then by function....components, so we will discuss them together. Gene expression The reason for most viral experiments is to deliver... deliver a genetic cargo, typically an expression construct, to cells/organisms. This is often cDNA, shRNA...

  2. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    Published
    May 13, 2025, 1:15 p.m.
    ...lentiviral vectors, how they work, and why you might choose them over integrating lentiviral vectors.  Before...integrase-deficient lentiviral vectors, let’s briefly revisit the basic biology of lentiviral vectors (for a more detailed...lentiviral vectors that are competent for one round of infection (AKA transduction in the viral vector world...life cycle of a lentiviral vector with an integrase-deficient viral vector. Created with BioRender.com...leading to transient expression of gene products (Wu et al., 2004). Over time, the expression diminishes as ...lentiviral vectors? There are several reasons why you might choose an IDLV over a lentiviral vector: 1. No...2008). 2. Transient expression of transgenes/shRNAs If you want your cells to express a protein or nucleic...

  3. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ... handling and expression of these vectors when used in experiments. Retroviral vectors make up a large...Transfer Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning...cloning and expressing your gene of interest with GFP expression.  Cloning and expressing RNAi pMKO.1 ...interest. Lentiviral vectors Popular Lentiviral Transfer Vectors    Expressing your gene of interest ...lentiviral vector backbone from the Shih lab. You can find more lentiviral expression vectors (empty backbones...building block Entry vectors and 5 lentiviral Destination vectors. These vectors can induce combinatorial...thereby enable long-term stable gene expression. Gamma-retroviral vectors are derived from the Moloney Murine...

  4. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    Published
    Aug. 19, 2014, 3:39 p.m.
    ...fusion protein. Mammalian vectors One of our most popular mammalian expression vectors, c-Flag pcDNA3, comes...most cases bacterial expression vectors are inducible (e.g. pBAD LIC cloning vector (8A)) and have an epitope...folding assays. Yeast vectors There are several types of yeast expression vectors. Consider your applications...packaging vectors to generate the virus. Bacterial vectors This system is great for overexpressing YGOI in...is your expression system or environment. The host organism will determine the type of vector that you...as close as possible to the basal expression level, induce expression at an opportune time or go all out...transforming wild-type yeast with an integrating vector expressing a selection cassette with 5′ and 3′ flanking...

  5. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...with AAV DIO/Flex vectors and leaky expression can occur in cells that don’t express Cre or Flp, what ...using DIO/FLEx vectors for off-target expression.  Some classes of neurons transiently express TH during development... require gene expression in a specific cell type rely on transgenic models that express recombinases like...and recombinase-dependent AAV vectors for selective transgene expression. While this is a powerful experimental... is off-target or spurious expression of recombinase-dependent AAV vectors. In this article, we will discuss...off-target expression and ways to optimize experiments to minimize it. Overview of DIO/FLEx vector design ...strategy should work, these vectors suffered from high levels of spurious expression.  As such, researchers...

  6. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ... of Gateway cloning vectors Vector type  Vector features Purpose Donor vector attP sites for recombination...Gateway vector to produce the desired expression clone. This is possible thanks to the Gateway vectors’ design...sequences into multiple vector systems. With the appropriate entry and destination vectors, one can use Gateway...reaction creates an expression clone with all of the components necessary for gene expression.     The ...destination vector. This reaction is catalyzed by the LR Clonase enzyme mix. As a result, an expression clone...ccdB gene is present in the donor vectors and the destination vectors prior to recombination, and it is...donor vector.   Method B: TOPO-cloning of the desired insert into an attL-entry-TOPO vector. TOPO...

  7. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...ideal split vector system. Overall the efficiency of transgene expression from split vectors has been inconsistent... overviews of three split vector strategies plus one additional split vector strategy for increasing the...homology directed repairs (HDR). Split AAV vector approaches Split vector approaches increase the size of the...Generally an overlapping split vector has equal or higher transgene expression when compared to using a single...copies of both vectors. Examples of use: Halbert et al., 2002 used an overlapping vector with 440 or 1000bp...dystrophy. While mini-dystrophin expression levels were lower than expression of wild-type dystrophin in normal...to deliver LacZ to muscle in mice. Expression from the hybrid vector, as measured by β-galactosidase activity...

  8. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ... for each gene that is expressed, multicistronic vectors simultaneously express two or more separate proteins...cells that are expressing your gene of interest at high levels. Unlike vectors that express screenable or...them. Why Use multicistronic vectors? Detecting cells that are expressing your gene, especially if you...want to express multiple genes in the same cell. Although it is possible to drive co-expression by using... with multiple, individual expression cassettes, having the genes expressed from the same cassette is ...relative expression levels between two or more genes is important. How do multicistronic vectors work? Scientists...transfer vectors with limited packaging capacity. Additionally, it may not be feasible to express more than...

  9. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...(tTA, also expressed by your transfer vector) binds to the TRE and activates the expression of your gene...your experiment using the lentiviral vector system. Viral vectors have been increasingly popular in fundamental...primary cells. Amongst the different vectors used, lentiviral vector constructs have proven very useful ...fluorescent sensors. Retroviral vector background Until the development of viral vectors, gene transfer into mammalian...un-transfectable cells, labs use viral vectors and particularly retroviral vectors because they target a broader...attractive choice for a gene transfer vector. Lentiviral vectors Although retroviruses are still in use...generation of lentiviral vectors consisted of three plasmids encoding 1) the lentiviral vector genome which was...

  10. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    Published
    April 28, 2016, 2:30 p.m.
    ...elements to turn off expression of one gene, while simultaneously turning on the expression of another! What...FLEx switch that turns BFP expression off, while turning on mCherry expression. For this FLEx switch to ...allowing for expression of GeneR. This allows a researcher to test whether or not expressing GeneR alone...potential to express the transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic...ensures Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...the presence of Cre, GFP expression is lost, allowing for rescued expression of GeneR.   Sometimes...Genome Engineering Using FLEx AAV Vectors? Learn about Viral Vector Elements Read Our Primer on Optogenetics...

  11. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...lentiviral expression of OSKM reprogramming factors: Mouse Expression Vectors Human Expression Vectors... residual expression piggyBac Excisible Medium Medium Generates transgene-free and vector-free cells...harmful viral particles that express oncogenes, such as Myc. Viral vectors also have the largest genomic...so either a single polycistronic vector or multiple single-gene vectors can be used to package reprogramming...reprogramming. Additionally, lingering expression or re-expression of viral transgenes in iPSC-derived cells... retroviral vectors. This makes it possible to use a polycistronic cassette that expresses multiple reprogramming...reprogramming vectors available from Addgene. Another drawback is that lentiviral vectors insert in different...

  12. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ...(scAAV) vector. Full-length ssAAV vector genome of both polarities are rescued from the vector plasmid...re interested in expressing multiple small genes, you can design single rAAV vectors with multiple genes... to co-express them from one promoter. Specificity The ability of AAV to drive gene expression in specific...cell-specific gene expression and viral infection. Cre or Flp dependent switches control gene expression via switches...transgene expression, since you don’t have to wait for the host to replicate the DNA before expression. Now..., and Constance Cepko. "Distinct expression patterns of AAV8 vectors with broadly active promoters from...about Viral Vector Elements Resources on Addgene.org Read Our AAV Guide Find AAV Vectors Browse All ...

  13. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    Published
    Sept. 29, 2015, 2:30 p.m.
    ...Agrobacterium binary vectors Vectors designed for use in Agrobacterium are termed binary vectors because they ...novel versatile gene expression vector for transient assays as well as stable expression of foreign genes ... and versatile binary Ti vector for both transient and stable gene expression applications in plants. ...pSIM24! To build a better binary vector, Sahoo et al. started with a vector commonly used by their lab, the.... This smaller, easily customizable vector functions in multiple species, including tobacco and Arabidopsis...coli. Like plasmids used in other systems, binary vectors are cloned and prepped in E. coli. However, they...co-cultivation with the target plant species. Thus, binary vectors contain two origins of replication, one each for...

  14. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...sequence into distinct expression vectors using the enzyme BbsI. These expression vectors each contain Type...Lentiviral Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing wt humanized...containing vector using Golden Gate assembly, and up to 8 gRNAs may be expressed simultaneously. Vectors for ...for ordered assembly into a destination vector that expresses Cas9. This is illustrated in the schematic...Cas9 or dCas9-containing destination vectors. Destination vector options include humanized wt Cas9, dCas9...Each destination vector contains GFP, enabling you to select cells with high GFP expression. These cells ...enables users to express up to 7 gRNAs. The kit contains different destination vectors depending on the...

  15. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    Published
    May 9, 2017, 2:12 p.m.
    ...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...

  16. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    Published
    Feb. 16, 2023, 7:05 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ... 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...

  18. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ... 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...

  19. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    Published
    March 25, 2014, 3:15 p.m.
    ...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...

  20. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    Published
    May 20, 2025, 1:15 p.m.
    ... 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...
Showing: 1 - 20 of 294 results