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We narrowed to 407 results for: expression vector

Showing: 1 - 20 of 407 results

  1. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...stable cell lines expressing a gene of interest from an integrated lentiviral vector....stable cell lines expressing a gene of interest from an integrated lentiviral vector. Unlike the short-term...using lentiviral vectors enables long-term protein expression studies. Moreover, repeating experiments ... the most desirable level of expression. Over time, transgene expression in a polyclonal population may...Lines with Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...short-term protein expression observed using transient transfection approaches, generating cell lines using...repeated transient transfection. Some lentiviral vectors deliver mammalian antibiotic resistance (e.g., ...

  2. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...bacterial expression vector. Using subcloning, you can easily move YGOI into a mammalian expression vector...same orientation on your target vector. If you are not sure what vector to use, you can check out our Empty...Congratulations, you now have YGOI in a mammalian expression vector and can begin your studies. Reference Page...be used to easily move any piece of DNA from one vector to another as long as it is already bounded by ...of interest (YGOI for short). You might need to express YGOI in cultured mammalian cells. The problem is...orientation in the recipient plasmid. (You don't want to express the antisense version of your gene!) Are in frame...manufacturer’s instructions. Isolate your insert and vector by gel purification Run your digested DNA on an...

  3. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning vector. The vector is 8901 base...-Cloning Vector I.2 Recipes I.3 Warranty information Back to Top A. pLKO.1-TRC Cloning Vector A.1 The ...use of this vector. A.2 Map of pLKO.1 pLKO.1 is a replication-incompetent lentiviral vector chosen by the...TRC-cloning vector. Addgene Plasmid Description pLKO.1 – TRC control Negative control vector containing... pLKO derived vectors that may also be useful for your experiment. To see these vectors, visit Addgene...and ligated into the pLKO.1 vector, producing a final plasmid that expresses the shRNA of interest. C.1...TRC Cloning Vector The following steps were the prevailing guidelines when the pLKO.1 vector was created...

  4. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...could lead to reduced transgene expression over time, as the lower expressing clones take over the polyclonal...by Limiting Dilution You may also like... Viral Vector Guides Virus Blog Posts Addgene Protocols Viral...that are more likely to retain stable transgene expression. Other methods of generating a monoclonal cell...population be monitored regularly for transgene expression. This protocol has been optimized for adherent...sufficiently expanded, screen the lines for transgene expression and/or other phenotypes. For example, perform...for lines with the highest or lowest transgene expression ( Figure 2 ). Sample Data Figure 1: Generation...from expansion of individual A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9...

  5. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...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...

  6. General Transfection

    Type
    Protocol
    ...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...polyethylenimine. Transfections allow for transient expression of a gene of interest in a target cell line and...the cells 24–48 h before checking for protein expression. Sample Data Legend: Lenti-X 293T cells were ...here by the amount of green fluorescent protein expression (green in the right panels) with a limited effect...

  7. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...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...

  8. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ... 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...

  9. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  11. Fluorescence Titering Assay

    Type
    Protocol
    ...Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...marker. Determining the titer of your lentiviral vector allows you to control the multiplicity of infection...protocol was developed using low titer lentiviral vectors (10 5 TU/mL). If you anticipate that your viral... transduction, cells were assayed for mCherry expression using fluorescence microscopy. Last reviewed ...

  12. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ... 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...

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

    Type
    Blog Post
    ...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...

  14. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ... 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...

  15. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...(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...

  16. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...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...

  17. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...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...

  18. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...(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 ...

  19. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    ...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...

  20. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...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...
Showing: 1 - 20 of 407 results