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We narrowed to 370 results for: Cre expression vector

Showing: 41 - 80 of 370 results

  1. Using AAV for Neuronal Tracing

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    Blog Post
    ... made Cre-dependent. If Cre-dependent vectors are used in transgenic animals that express Cre in only ...of infected cells that also express Cre. Another advantage of AAV vectors is that there are well-established...et al. 2014 traced neurons with an AAV9 vector expressing a Cre-dependent fluorescent marker. Injection...communication. Viral vectors and the nervous system With the advent of viral vector technology, new methods... at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have been widely used...transgene expression. Fluorescent proteins are widely used for tracing and their expression is often made... into a conditional mutant mouse brain that expressed Cre recombinase only in pyramidal cells of the hippocampal...

  2. Viral Vectors 101: Systemic Capsids

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    Blog Post
    ... be paired with Cre-expressing transgenic mouse lines to restrict transgene expression to specific tissues...recall, are the outer shell of an AAV vector, which determine the vector’s tropism, including its ability to...used to prevent expression in cell types not of interest. Systemic capsids packaging Cre-dependent transgenes...all take up precious cargo space in AAV vectors. Dual expression systems, which use two or more AAVs to...payloads. Since most systemic vectors are primarily derived from AAV9, the vectors have to be administered ...on the Addgene blog Viral Vectors 101: AAV Variables That Matter Viral Vectors 101: An Introduction to ...Selection of an Efficient AAV Vector for Robust CNS Transgene Expression. Molecular Therapy - Methods ...

  3. Viral Vectors 101: Calcium Sensors

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    Blog Post
    ...mammalian cell expression, and compatible with constitutive or Cre-dependent expression. These sensors...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...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 ...

  4. Viral Production at Addgene

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    Blog Post
    ... the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core. Transfections...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....Transducibility Some viral vectors are tested in vitro and in vivo for gene expression and/or function. These....1E6 vg/mL, or both. Two days later, Cre-dependent GFP expression was detected with direct fluorescence...Empty vector particles can be identified after negative staining and appear darker than full vector particles...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...

  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...post transfection when creating stable cell lines. Looking for Mammalian Expression Plasmids? Check Out...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...

  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...Figure 1: Shuttle vectors are vectors that can function in more than one host species. Created with BioRender.com...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. Viral Vectors 101: Viral Applications

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    Blog Post
    ...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. ...immunity. Viral vectors are being explored as the delivery mechanism for increasingly popular nucleic ...

  8. 15 Years of Addgene: The Top 15 Plasmids

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    Blog Post
    ...  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...

  9. Fluorescent Tagging of Endogenous Genes with SapTrap

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    Blog Post
    ...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...

  10. New and Upcoming Viral Vectors - June 2019

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    Blog Post
    ...with Cre for intersectional control of specific neuronal populations. We now offer a Flp-expressing AAV...AAV, as well as a Cre-dependent Flp-expressing AAV. See our recombinase AAV inventory here New recombinases...AAV1 pAAV-ihSyn1-DIO-tTA 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...new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV we have released in the...detection PSAM4 GlyR for chemogenetic inhibition Flp expression for intersectional studies And more! Control...Recombinase AAV Finally, we are excited to add Flp-expression AAV to our recombinase AAV inventory. Flp is ...

  11. Lentiviral Vector Uses and Overview

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    Blog Post
    ...Plasmid FAQs. Lentiviral vectors in the lab Lentiviral vectors are incredibly popular in the lab - the...and genome-wide screening.   Lentiviral vectors in the clinic Since lentiviral vectors can deliver a large...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 ...

  12. Viral Vectors 101: AAV Variables That Matter

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    Blog Post
    ...early gene expression, but can become methylated over time, resulting in decreased expression (Damdindorj...know what expression pattern you need, and the factors in your system that can affect expression, before...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...

  13. 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...and rAAV vectors, modifications can affect gene transfer efficiency and transgene expression (Zhou et ...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...reduce the packaging efficiency but increase transgene expression (Zhou et al., 2017). The most common...transcription factor binding sites can increase transgene expression and transduction efficiency (Shitik ...repeats reduces rAAV productivity but increases transgene expression. Sci Rep, 7(1), 5432. https://doi.org...

  14. 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...

  15. 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...

  16. An Introduction to Adenovirus

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    Blog Post
    ...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...

  17. Plasmids 101: Control Plasmids

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    Blog Post
    ...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... the decreased expression of Gene X. Ergo the positive control(s) should decrease the expression of Gene...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...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 ...

  18. Plasmids 101: Blue-white Screening

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    Blog Post
    ...operon repressor, thereby allowing lac expression). When expressed, the β-galactosidase enzyme can break... methods. Positive selection vectors encode a gene which, when expressed, is lethal to the cell. Cloning...other screening strategies, check out our screening overview blog post.) *Note: Two Popular Vectors for...pBluescript are a few common vectors. Blue-white screening is just that – a screening process. It does not select... inserted it into a plasmid, creating an α-complementation cloning vector. When a cloning reaction goes... that encodes for the enzyme β-galactosidase. Expression of the lac operon is induced by lactose, and ...enough time for any intact β-galactosidase to be expressed and process x-gal into blue pigment (16-20 hours...

  19. Sleeping Beauty Awakens for Genome Engineering

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    Blog Post
    ... of vectors for the SB transposase system - one set of vectors for robust constitutive expression (pSBbi...gene (red) for expressing the transposase enzyme.2.) The transposase enzyme is expressed (red stars) and...efficiencies of integrating viral vectors. New Sleeping Beauty transposase vectors Even more recently, Eric ...pSBbi derivatives) and one set of vectors for inducible expression (pSBtet derivatives). The constitutive...constitutive and inducible expression vectors were developed into 16 variants with different selection markers...cells’ defense mechanisms. The use of non-viral vectors, like transposons, avoids many, though not all,...transposition; and 2) a transposon containing the gene-expression cassette that can translocate within the genome...

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

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    Blog Post
    ...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...tracked, and appropriately credited, broadening the knowledge base of AAV vector performance in various brain...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 ...

  21. New and Upcoming Viral Vectors - Spring 2019

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    ...restricts expression to GABAergic interneurons. These vectors can also be used as the control vectors for experiments...and control AAV These fluorescent protein expression vectors can be used to visualize cells and to quantify...pAAV-hSyn-mCherry at Addgene. Viral vectors coming soon! These vectors listed below should be released at...DreO-bGHpA 121675  AAV9  pAAV-EF1a-fDIO-Cre (FLP-dependent CRE) 55634  AAV1  pAAV-EF1a-mCherry-IRES-... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV we have released so far... constitutive (pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-...

  22. Synthetic promoter AAVs for cell-type specific expression in retinal cells

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    Blog Post
    ...promoter to drive expression of Cre-mCherry and a second promoter to drive expression of a Cre-dependent fluorescent...reporter. Expression of GFP will only result in cells where Cre-mCherry is also expressed. If Cre-mCherry...Cre-mCherry expresses in photoreceptors and horizontal cells, and Cre-dependent-GFP expresses in horizontal cells...to test for cell type-specific expression of GFP. Half (113) expressed in retinal cells four weeks post-injection...More than 90% of expression occurred in one type of cell More than 50% of expression occurred in one type...of targeted expression. Surprisingly, <1% of synthetic promoters replicated the expression specificity...restrict expression to a single cell type based on the overlap in cell types both promoters expressed in. ...

  23. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

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    Blog Post
    ... an episomal vector system that increases reprogramming efficiency through the expression of a shRNA against...cells, thus expressing additional EBNA-1 transiently. The transiently expressed EBNA-1 increases protein ...RNA virus Episomal vectors- extrachromosomally maintained DNA plasmids that express reprogramming factors...Reprogramming efficiency of PBMCs with episomal vectors Episomal vectors are based on the Epstein-Barr Nuclear ...long-term expression of the reprogramming factors. However, reprogramming of PBMCs by episomal vectors has ...protein expression from the other episomal plasmids. When used on PBMCs, the pCXLE episomal vector system increases...method for creating clinical-grade, safe, nonviral and nonintegrating hiPSCs where the vectors encoding ...

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

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    Blog Post
    ...variety of pExpression vectors) plasmids. The type IIS restriction sites used in these vectors are of low...Confronted with the problem of creating vectors for predictable protein expression in bacteria, researchers ...SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of proteins...or a mammalian expression vector in a one-pot reaction. Using this system, you can create custom RNA binding...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, ...Process with pCri Cell-free protein expression kit Cell-free expression systems have been improved for both...

  25. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

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    Blog Post
    ... tool for gene expression in mammals. Resources on Addgene.org Search for AAV vectors to use with your...pathogens to the CNS, but this also stops viral vectors, such as adeno-associated virus (AAV), from reaching...injection into the bloodstream, but requires a high vector dose and is unable to reach all regions of the ...phenotypic effects such as those that affected gene expression, mRNA splicing, or protein coding regions. In...one, Ly6a, showed differential protein and RNA expression between the brains of C57BL/6J mice and BALB/...strong linkage to the PHP.B phenotype and its expression pattern in the mouse brain.  Both labs then took...Deverman lab started by seeing how changes in LY6A’s expression affected PHP.B’s transduction rates. Knock out...

  26. Plasmids 101: Protein Expression

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    ...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...

  27. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

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    ... expression. CAG-FLEX: CAG promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin...neuronal expression. hSynap-FLEX: Synapsin-1 promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP...systems you can use to express the iGluSnFR3 variants: pRSET: Bacterial expression vector, use with IPTG induction...transmembrane domain in the mammalian expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol...FLP-dependent expression GFAP: Glial fibrillary acidic protein (promoter). Good for Glial expression (not neurons... eGFP for sfGFP to create SF-iGluSnFR. This improved variant has higher expression levels in bacteria ...range, photostability, quantum yield, and expression. Create yellow (v82) and lime green (v857) variants...

  28. Rabies and Neuronal Tracing

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    Blog Post
    ...Control Gene Expression with FLEx Vectors Resources on Addgene.org AAV Vector for TVA Expression AAV for ...example, a researcher can deliver Cre-dependent TVA to Cre-expressing neurons in a transgenic mouse line...RABV expressing the viral protein EnvA on its surface will only infect Cre-positive, TVA-expressing cells...Addgene Blog AAVs CREATed for Gene Delivery to the CNS and PNS Advanced Uses of Cre-lox and Flp-FRT - ...to visualize how neurons are connected. Viral Vectors as Neuronal Tracers Due to their ability to efficiently...achieved by using EnvA-pseudotyped RABV, and only expressing the EnvA receptor (TVA) in user-defined cells... genome, so viral progeny will not necessarily express EnvA unless it is supplied in trans. Monosynaptic...

  29. Easing the Protein Purification Process with pCri

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    Blog Post
    ... information, you can then pick an expression vector and expression system. If you find that your protein... al. "The pCri system: a vector collection for recombinant protein expression and purification." PloS ...protein might not fold well, you should screen a few different vectors to see which gives you the most functional... the secretory pathway to improve folding and disulfide bond formation. If one prefers expression in E...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...pCri also includes plasmids designed for protein expression in systems other than E. coli, specifically the...

  30. Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering

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    ...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...

  31. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

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    ...Learn about Using Cre in Drosophila Read Other Genome Engineering Posts Use FLEx Vectors for Site-Specific... plasmids for overexpression and knockdown. Addgene also has a number of insect vectors to consider. An... your gene of interest in a plasmid and get the vector incorporated into the germline. To begin this process... browse Addgene’s Collection under the insect expression system or by filtering by fly as species of gene...Find Gal4 and UAS plasmids at Addgene Choosing a vector to generate a transgenic fly There are a few important...important points to consider when choosing the vector you’d like to use to generate your new fly line:... gene to be incorporated into the genome? Some vectors incorporate into the genome randomly, while others...

  32. Viral Vectors 101: Pseudotyping

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    ...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 ... Lentiviral vectors pseudotyped with a modified RD114 envelope glycoprotein show increased stability in...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...

  33. Hot Plasmids and Viral Preps - May 2021

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    ...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...

  34. 28 Hot Plasmid Technologies from 2015

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    ...bacterial expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT...luciferase) to create a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed... to create the pCell-Free vectors, Gateway-compatible backbones which enable cell-free expression of proteins...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ... SFFV) or inducible (CMV-TO) cDNA expression, sgRNA (U6) expression, shRNA/shRNA-miR30 constitutive (H1...-miR30 (CMV-TO) expression, or a promoterless construct. Pre-constructed Entry vectors containing Cas9...reaction with a desired destination vector expressing a separate reporter gene (EGFP, iRFP, IFP1.4, puromycin...

  35. AAVs in Retinal Gene Therapy

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    .... 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 ...

  36. 27 Hot Plasmids from 2016

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    ...AAV transfer vector for in-vivo expression and plasmids optimized for dCas9-VPR expression in drosophila...parasites that constitutively express Cas9 with a library of sgRNA expression vectors (containing ten guides ...easily create protein expression constructs fused with epitopes or fluorophores, generate expression vectors...large-scale expression screen that can be used to generate His-tagged human kinase constructs that express well... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ... to test multiple expression systems before they achieve the appropriate expression level for their gene...various promoters suitable for in vivo expression, and/or produce vectors to make mutant cell lines or transgenic...

  37. AAV Vector Quality Control: Going the Extra Mile with NGS

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    ...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...producing multiple vectors in parallel because the risk of accidental errors increases. In addition to human... 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...

  38. Your Top Requested Plasmid in 2016!

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    Blog Post
    ...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...

  39. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

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    ... 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...

  40. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

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    ...multicolor labeling, Brainbow used the Cre-loxP system to express one of the three fluorescent protein ...strategies. In such vector-based gene delivery systems, the stochastic expression of XFPs is possible ...reduced expression level of XFPs, most likely by suppressing transcription. It is critical to express a minimal... stochasticity will result in a combinatorial expression of these three genes with different copy numbers...This is because the stochastic and combinatorial expression of fluorescent proteins is only possible at low...Tetbow In modern neuroscience, plasmid or viral vector-mediated gene delivery is more common than conventional...Poisson distribution. To overcome the limited expression levels of XFPs in the original method, we used...
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