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We narrowed to 370 results for: Cre expression vector

Showing: 1 - 40 of 370 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...is described in Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, ...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...

  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 The following steps were the prevailing guidelines when the pLKO.1 vector was created. For...-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...

  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...have been sufficiently expanded, screen the lines for transgene expression and/or other phenotypes. For example...Western blotting to screen for lines with the highest or lowest transgene expression ( Figure 2 ). Sample...is described in Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, ...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...

  5. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...to use the pLKO.1 vector can be found here. Cre-Lox technology Puro.Cre empty vector (Plasmid #17408)   ... 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 ...Tools for RNAi! Cre-Lox technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage...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...

  6. 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 can...can be expressed in cells where Cre is absent. Additionally, we have experienced that some vectors are much...using DIO/FLEx vectors for off-target expression.  Some classes of neurons transiently express TH during development...off-target expression. Figure created with BioRender.com   Strategies for optimizing expression So now... require gene expression in a specific cell type rely on transgenic models that express recombinases like... like Cre or Flp in their cells of interest and recombinase-dependent AAV vectors for selective transgene... is off-target or spurious expression of recombinase-dependent AAV vectors. In this article, we will discuss...

  7. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...have created a cassette that will express GeneR only in the presence of Cre. In this example, Cre would...tissue-specific Cre is provided. Since the FLEx switch ensures Cre-dependent reporter expression, the potential...: In the presence of Cre, GFP expression is lost, allowing for rescued expression of GeneR.   Sometimes...elements to turn off expression of one gene, while simultaneously turning on the expression of another! What...SSRs) Cre and FLP. SSRs bind DNA at target sites to induce site specific recombination events: Cre recombinase...FLEx switch that turns BFP expression off, while turning on mCherry expression. For this FLEx switch to ...Basic FLEx switch will express BFP in the absence of, or mCherry in the presence of Cre recombinase.   ...

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

    Type
    Blog Post
    ...inject your Cre-dependent mCherry mouse with the pmSyn-EBFP-Cre AAV. Because Cre expression is controlled... can be used to drive Cre expression specifically in neurons. AAV Delivered Cre - A useful tool for neuroscientists... could breed your Cre-dependent mCherry line with a separate line that expresses Cre in neurons, but both...pmSyn1-EBFP-Cre and other viral vector tools in the years to come. We’ll make many more viral vector tools ...most popular form), this viral vector is used to deliver DNA encoding Cre recombinase under the control...control where your gene of interest is expressed.  A cell-type specific Cre can provide you with that control...with a means to specifically activate expression of any Cre-dependent gene of interest in neurons. This ...

  9. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ... cycle of a lentiviral vector with an integrase-deficient viral vector. Created with BioRender.com. ...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...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...

  10. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...transfected with both vectors. Flex vectors are the solution. Flex vectors expressing fluorescent cytoplasmic...control FLP and Cre expression have been developed. Read on to learn more about FLP-FRT, Cre-lox, and how... vivo, FLEx vectors expressing the light activatable ion channel under the control of Cre recombinase ...concentrations of vectors expressing the required recombinase. The high concentration of FLEx vectors guarantees...LDB) to the C-terminus of FLP or Cre (pCAG-CreERT2 #14797). These Cre/ FLP versions are retained in the...control without modification - either Cre is there or it’s not. Cre-mediated possibilities for site specific...sequence) and expressing it from a CAGGs promoter. However FLPe is still less efficient than Cre recombinase...

  11. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    ...the Esr1 domain that blocks Cre expression. Then, Cre is expressed and Cre-mediated recombination of floxed...that regulates Cre expression. The most common drug used to induce the expression of Cre is tamoxifen. .... In tamoxifen-inducible Cre (Cre-ER) systems, Cre is fused to the ligand binding domain of estrogen receptor...doxycycline mediated activation of Cre to either induce or repress Cre expression using TetON or TetOFF systems...or eliminates the expression of a particular gene. Targeting vectors used to create knockout mice include...targeting vector, creating the floxed gene segment. Crossing of mice containing a floxed gene to a Cre-expressing...knockout/knockin mice, use the Cre-lox recombination system to express the transgene in specific cells...

  12. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...permanently induce expression? Check out our Retro expression vector and Lenti expression vector. Turn it on...protein expression. If you need the ability to control when you turn off expression for good, the Cre-Lox...knock out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for a little while ...induce expression? Check out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off...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...

  13. New and Upcoming Viral Vectors - September 2019

    Type
    Blog Post
    ...Flp-dependent Cre AAV in AAV9 and AAVrg. See our new additions here: Cre vectors pAAV-EF1a-fDIO-Cre (121675...directs strong, ubiquitous expression. This is a Cre-dependent switch that expresses nuclear mCherry in the...the absence of Cre, and expresses nuclear EGFP in the presence of Cre. Biosensor AAV (calcium sensors ...ubiquitous expression. pAAV-mDlx-NLS-mRuby2 (99130-AAVrg): The Dlx element directs expression in forebrain... Flp vectors pAAV-EF1a-mCherry-IRES-Flpo (55634-AAV1) pAAV-EF1a-Flpo (55637-AAV1) Viral vectors coming... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...121675-AAV9, 121675-AAVrg)  pENN.AAV.hSyn.Cre.WPRE.hGH (105553-AAV5) Dre vector AAV phSyn1(S)-DreO-bGHpA...

  14. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...cell-specific gene expression and viral infection. Cre or Flp dependent switches control gene expression via switches...(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...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 ...

  15. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...that express cre recombinase. Typically, this feature is used to attain cell-specific expression in transgenic...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....lentiviruses expressing various gRNAs. It’s better to compare the phenotypic effects of expressing various ...transductions with a single-vector system). Tips for using this plasmid Cas9 expression is typically cytotoxic...

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

    Type
    Blog Post
    ...cell-type-specific Cre, the polyA sequence is inverted only in the Cre-expressing tissues. This inversion creates a template.... By using this library to infect an animal expressing Cre in a cell type of interest, a scientist can...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...studying the CNS and PNS! What is the CREATE method?  CREATE stands for Cre Recombinase-based AAV Targeted Evolution...

  17. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...lentiviral expression of OSKM reprogramming factors: Mouse Expression Vectors Human Expression Vectors...Excisable Cre-lox lentiviral vectors: This approach is the same as using a standard lentiviral vector, but ... 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...

  18. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...from Cre-based Brainbow-expressing cells. Autobow plasmids are all-in-one versions which express self-... existing transposon and vector systems and creating an all-synthetic vector that included only the elements...compatible Level 0 vectors are directionally assembled into a Level 1 vector creating a single transcriptional... 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.,... These vectors, termed RUSH (For ROSA26 U6 short hairpin) and CRUSH (Conditional RUSH) use Cre-mediated...

  19. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    ...preps for expressing SARS-CoV-2 proteins ChR2 for targeted expression DREADDs GFP dependent Cre and FLPo...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...serotypes, as well as a new Cre dependent GFP control plasmid for expression in forebrain GABA-ergic interneurons...for targeted expression. Plasmid 75470 is now available in AAV1 for Flp-dependent expression of ChR2(H134R...dependent Cre. The two segments come together in the presence of GFP to form a functional Cre enzyme that...GFP expression patterns, thus allowing you to further restrict gene activation to cells expressing both...

  20. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...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...cases, AAV vector screens may be AAVantageous!       In vivo, in vitro, ex vivo CRISPR screens can be carried...preferred delivery method for in vivo CRISPR screens. AAV vectors can be made at high titers equal to those...Cpf1). Even more creative options involve generating strains of mice that stably express Cas effectors in... aided by advancements in in vivo CRISPR screens. AAV vectors have played a large role in these advancements...

  21. 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 ... remove cells and debris. Lentiviral titer can decrease during cycles of freeze-thaw. If you are freezing...

  22. Viral Vectors 101: The AAV Retrograde Serotype

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    Blog Post
    ...by injecting a Cre-dependent AAV into Broca’s area of a mouse specifically expressing Cre in the hippocampus...your system. Chemogenetic vectors  Expression of retrograde serotype vectors is present at both the injection...phenomenon is to use a retrograde recombinase-expressing vector (e.g. Cre or FLP) to transduce the projection pathway... However, the Cre-dependent AAV cargo would only become activated in the presence of Cre recombinase, ...-dependent vectors, this can cause challenges for the experimental design as these vectors can exhibit...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 ...

  23. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ... or by mating with Cre-expressing mice. Figure 1: (A) Schematic of the Cre-dependent SPH construct... (i.e., how many sgRNA expression cassettes will fit). Cre-dependent expression of the SPH activator paired... ubiquitous expression of sgRNA(s) and mCherry (vector 1) with neuron-specific expression of dCas9-VPR... of Cre-dependent mouse lines for overexpressing genes within discrete cell populations makes Cre-dependent...plasmids used to express CRE under neuron-specific promoters (aCaMKII and hSyn). Created with BioRender....ubiquitous expression of sgRNA (top) and neuron-specific expression of dCas9-VPR (bottom). Created with BioRender.com...construct). More importantly, the expression of this construct is Cre-dependent, enabling spatiotemporal...

  24. 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...manipulation can also reduce expression levels of certain genes. A popular method uses Cre recombinase on transgenic.... 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...

  25. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...knockdown, Tet and Cre-regulated expression, CRISPR genome editing, and more. Not surprisingly, we receive...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...

  26. Deciphering the Mysteries of Behavior with Viral Vectors

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    Blog Post
    ...researchers injected Cre-expressing AAV-5 into the DRN of a transgenic mouse harbouring a Cre-dependent IL-1... particles into a Drd1-Cre transgenic mouse to deliver Cre-dependent expression of  Channelrhodopsin 2...-1 beta overexpression cassette. Expression of Cre in these neurons turns on overexpression of IL-1 beta.... 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...neuroscience. With the ability of viral vectors to fine tune gene expression temporally and spatially, exciting ...all of our viral vectors blog posts Find tips for getting started with viral vectors for behavioral neuroscience...

  27. Viral Vectors 101: Parts of the AAV Transfer Plasmid

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    Blog Post
    ... of expression. For example, CAG is a strong promoter that drives very high, ubiquitous expression, while... with the Cre recombinase, are used to spatially and temporally control transgene expression. Lox sites...DIO. Lox sites are recognized by the recombinase Cre. Cre first flips the sequence between the first pair...aids in nuclear export of mRNA and helps increase the expression of your transgene. The WPRE is located ...outside of the ITRs does not get packaged in the AAV vector. The cloning capacity between the ITRs is ~4.4 ...of interest. Promoter The promoter drives the expression of the AAV transgene. On a plasmid map, promoters...type-specific, which can help you restrict the expression of your transgene to a specific species or location...

  28. Choosing Your Perfect Empty Backbone

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    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...wild-type yeast with an integrating vector expressing a selection cassette with 5′ and 3′ flanking sequences...

  29. Important Considerations When Using AAVs

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    Blog Post
    ... have found that expression occurs quickly and results in long and stable expression (up to 9 months in... a specific system for controlling gene expression such as Cre-lox, may be sufficient to overcome the ...the onset of gene expression. However, it has yet to be determined what gene expression looks like at 3 ...-associated viral vectors: packaging capacity and the role of rep proteins in vector purity." Human gene...Tool for Gene Expression in Mammals A Match Made in Heaven: CRISPR/Cas9 and AAV Viral Vector Elements Additional...days vs 7 days vs 14 days etc. To decrease the time until expression, it is possible to use AAV constructs...Future? I knew little about adeno-associated Viral Vectors (AAVs) before starting my graduate program at the...

  30. Hot Plasmids Spring 2024

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    Blog Post
    ... administering these AAVs with a dilute Cre-expressing vector resulted in a sparsely-labeled field of ... A cis-splicing vector served as positive control (cis-ctrl). v1, vector 1; v2, vector 2; BD, binding ... O’Neill New viral vector preps expressing the Voltron2 voltage indicator, created by the Schreiter Lab...SARS-CoV-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting microglia ...Ace2N-mNeon-A122D and Voltron2 are available as Cre-dependent AAV1 vectors and are suitable for extended imaging...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...

  31. Plasmids 101: Gateway Cloning

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    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...attR destination vector will recombine with the attL-entry clone to create the expression clone.    ...Generating an expression clone. The reaction between the entry clone and destination vector creates two products...sequences into multiple vector systems. With the appropriate entry and destination vectors, one can use Gateway...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...

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

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    Blog Post
    ...example Cre/loxP or Flp/FRT, with AAV vectors are common approaches to control transgene expression by restricting...restricting expression to genetically defined cell types. Specific transgene expression can be also achieved...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...

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

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    Blog Post
    ...ideal split vector system. Overall the efficiency of transgene expression from split vectors has been inconsistent...of three split vector strategies plus one additional split vector strategy for increasing the size of the... repairs (HDR). Split AAV vector approaches Split vector approaches increase the size of the gene delivered...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...

  34. New Viral Vectors - Fall 2024

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    Blog Post
    ...Armamentarium Collection. These preps drive expression of either Cre recombinase or a fluorescent marker in...AAV-GfaABC1D-Cre-4x6T AAV5 Recombinases Stanley Thomas Carmichael New viral prep pAAV-EF1a-fDIO-Cre AAV5 ...the 34 new preps available in the Addgene viral vector repository! This includes the first Addgene prep...X1.1 serotype.  In rodents, AAV9-X1.1 enables expression in brain endothelial cells via an injection into...actually targets neurons, and has 45-fold higher expression levels compared to AAV9. You can now target these... AAV9-X1.1!  We also have eight new AAV-PHP.eB vectors from the Jonathan Ting lab at the Allen Institute...carried out before. We have a bunch of other new AAV vectors as well — including next-generation biosensors ...

  35. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...optogenetic FLEx vectors in combination with transgenic animals or rAAVs expressing Cre under a specific...cells with an inactive opsin gene; in Cre expressing cells, however, Cre induces the recombination of the ...enable strong opsin expression in specific neuronal cells? This can be achieved using Cre-dependent viruses...FLEx-based vectors to study the neural circuitry behind the mechanisms of sleep and anesthesia. In Cre transgenic...reversible modulation of FLEx vectors combined with the use of several Cre transgenic mice targeting different...not naturally light-sensitive, so the selective expression of opsin genes in targeted cells makes it possible...How to get an opsin into the brain In order to express opsins within the brain, researchers inject genetically...

  36. Plasmids 101: Multicistronic Vectors

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    Blog Post
    ... for each gene that is expressed, multicistronic vectors simultaneously express two or more separate proteins...high levels. Unlike vectors that express screenable or selectable markers from a unique promoter, multicistronic...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...

  37. Quick Guide to All Things Lentivirus

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    Blog Post
    ...(tTA, also expressed by your transfer vector) binds to the TRE and activates the expression of your gene...experiment using the lentiviral vector system. Viral vectors have been increasingly popular in fundamental and...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...

  38. Plasmids 101: Knockout/Knock-In Plasmids

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    Blog Post
    ...Resources on Addgene.org Find Mammalian Expression Vectors Find Cre-Lox Plasmids Learn More about CRISPR ...: A knockout targeting vector designed to insert a resistance gene. The vector contains a neomycin resistance...the addition of Cre recombinase (Figure 2). Figure 2: A knockout targeting vector designed to insert...using Cre recombinase. If GFP is under control of an endogenous promoter, you can use expression GFP to...want to create a conditional knockout. To make a conditional knockout, researchers often use the Cre/Lox ...an intron.    Figure 4: A targeting vector to create a floxed allele for a conditional knockout...intronic regions, this gene will still be expressed. You'd first screen for this specific recombination outcome...

  39. 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 ...Maiti has created a new and versatile binary Ti vector for both transient and stable gene expression applications...pSIM24! To build a better binary vector, Sahoo et al. started with a vector commonly used by their lab, the...applications in plants. This smaller, easily customizable vector functions in multiple species, including tobacco...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...

  40. CRISPR 101: Multiplex Expression of gRNAs

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    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...
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