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We narrowed to 83 results for: Cre expression vector

Showing: 41 - 60 of 83 results
  1. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ... 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...
  2. Another Pathway into Cells: iTOP

    Type
    Blog Post
    Published
    June 23, 2015, 8:37 p.m.
    ...in which transient protein expression has a long-lasting effect, such as Cre- or Cas9-mediated recombination... mutations and gene expression changes elsewhere in the genome. Second, expression is by nature transient... Primary cells recapitulate the natural biology of a cell type of interest better than immortalized lines...control could enter cultured cells and activate expression of a luciferase reporter. CPP was not required...tested iTOP in a series of primary cell types using Cre and various loxP reporters. In both mouse and human...stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. CRISPR...potential application of iTOP Having tested iTOP using Cre-lox recombination, D’Astolfo et al. were eager to...
  3. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...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... 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...dual-color RNAi “sensor” mouse strain in which Cre expression causes the induction of both DsRed fluorescence...
  4. Rabies and Neuronal Tracing

    Type
    Blog Post
    Published
    May 29, 2018, 1:51 p.m.
    ...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...Uses of Cre-lox and Flp-FRT - A Neuroscientist's View Control Gene Expression with FLEx Vectors Resources...Addgene.org AAV Vector for TVA Expression AAV for Rabies Glycoprotein (G) expression  Find Retrograde...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...
  5. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ... 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...a conditional mutant mouse brain that expressed Cre recombinase only in pyramidal cells of the hippocampal...
  6. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    Published
    March 5, 2019, 1:08 p.m.
    ...promoter is expressed. Cre-mediated deletion of one or several of these genes permits the expression of genes...copy of the transgene expressing either YFP or CFP, and cells that do not express Cre fluorescing with the...activate the expression of a different fluorescent protein. Therefore, each cell could express a unique combination...protein genes, Cre recombination is required before fluorescence can occur. Cre recombinase can be used ...developing fish via Cre protein microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier...any cell type when crossed with mice expressing the appropriate Cre allele. The Confetti mouse contains...so that XFP are only expressed following a recombination event. Tissue-specific Cre recombination in this...
  7. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    Published
    Sept. 10, 2015, 2:30 p.m.
    ...with an appropriate mouse line expressing a particular cre, viral expression of your opsin is no longer required...steps in the coming weeks. Expression of the optogenetic constructs Expression of your desired optogenetic...promoter-directed expression alone is extremely limited. Therefore, highly expressing, generic neuronal...promoter of the cre recombinase while still taking advantage of the robust expression via the promoter...Finally, expression of the opsin in a mouse line will likely be less than that of viral expression but may...stimulation. Testing opsin expression  Regardless of the method of expression of your opsin gene, it is...extremely high expressing promoters). Conduct a time course experiment in which you monitor expression at various...
  8. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    Published
    April 7, 2022, 1:15 p.m.
    ... 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...SF-iGluSnFR. This improved variant has higher expression levels in bacteria and produces stronger fluorescent...dynamic range, photostability, quantum yield, and expression. Create yellow (v82) and lime green (v857) variants...
  9. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed in the cytosol...#140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly ...fluorescently labeled piggyBac vectors for uniform and sustained expression of single or multiplexed gRNAs...and track cells expressing both dCas9 and gRNAs. The multi-gRNA piggyBac vector integrates into ... the mCitrine gene. mCherry is constitutively expressed. Cells are then cultured and FACS sorted. Image...Histone 2A or 2B can result in bright targeted expression while minimizing non-specific labeling. For labeling...
  10. 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...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...
  11. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    Published
    Jan. 18, 2018, 2:34 p.m.
    ...inducible system is found in many bacterial expression vectors. Negative inducible promoter pBad is another...gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant...Temperature inducible promoters Temperature sensitive expression systems are typically less leaky than chemically...chemically induced promoters; they show near-zero expression at regular temperatures but can be induced by ... promoters, in which a gene of choice is only expressed following exposure to a brief heat shock. In the...Addgene depositors have developed heat shock-inducible Cre and Cas9 for easy genome engineering in species like...promoters Light is another way to activate gene expression, and two-component systems used in synthetic ...
  12. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ... minimize expression levels of your gene of interest, as needed. Controlling the expression levels of ...quickly transfer a DNA sequence from the transposon vector to one of many TTAA sequences distributed throughout...encourage you to deposit your piggyBac™ transposon vectors with us to help us expand this useful resource....may be particularly useful when the gene being expressed has threshold effects, such as with signaling ... can also mitigate the deleterious effects of expression of certain genes, such as those that have negative...piggyBac™, you could use recombinase technology (Cre/loxP) to remove the donor cassette. While this method...edit of interest. With the larger piggyBac™ donor vector, it is possible to make the ITR and selection cassette...
  13. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    Published
    March 21, 2019, 12:47 p.m.
    ...doxycycline, but it can also be regulated spatially with Cre-recombination. Read the paper in Frontiers in Molecular...single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one overnight...overnight reaction. STAgR is highly customizable as vector backbones, and gRNA structure, numbers, and promoters...Read the paper in Science Find the anti-CRISPR expression plasmids CRISPR-based method for cell lineage...Averof lab. It’s an alternative to the conventional Cre or Flp recombinases used to excise markers. It has...
  14. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    Published
    Dec. 7, 2015, 6 p.m.
    ...CRISPR genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate inversions, deletions...researchers with many different ways to control gene expression. Get the full scoop from Addgenie A. Max Juchheim... rundown of some of the strains you can use to express your protein of interest. 4. Plasmids 101: Blue-White...Blue-White Screening Getting a lot of empty vector contaminants in your cloning experiments and tired of...CRISPR/Cas9 and AAV Adeno-associated viral (AAV) vectors are useful for in vivo gene delivery, but have ...Browse Our CRISPR Resources Learn More about Viral Vectors Browse the Repository  ...
  15. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    Published
    Nov. 27, 2019, 2:03 p.m.
    ...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...
  16. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ... 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...Future? I knew little about adeno-associated Viral Vectors (AAVs) before starting my graduate program at the...in locomotion. We have relied heavily upon AAV vectors to deliver chemogenetic/optogenetic constructs ...
  17. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ...from other regions of the cell. By combining vectors expressing different fluorescent proteins targeting ...display (FingRs), Xue Han’s lab has developed AAV vectors to selectively visualize excitatory and inhibitory...excitatory and inhibitory synapses, respectively. Once expressed in the neuron, the FingRs bind to the endogenous...targeting inhibitory or excitatory synapses or that are Cre-inducible, researchers can explore synaptic integration.... The team engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other cellular...proliferation in acute myeloid leukemia and saw that expression of H2B-FT consistently predicted the cell cycle...different cell types. In one example, H2B-FT was expressed in murine B cells (BaF3), which depend on IL-3...
  18. Hot Plasmids: Fall 2025

    Type
    Blog Post
    Published
    Nov. 11, 2025, 2:14 p.m.
    ...cells express iCre with a mitochondrial anchor and a TEV cleavage site. Recipient cells express DsRed ...Once the recipient cells take up the Cre-tagged mitochondria, Cre is released, translocates to the nuclease...killswitch (GFP-nb–KS) is delivered using a vector that also expresses mCherry to mark transfected cells. The...cell-surface proteomes (Wu et al., 2025). The team expressed extracellularly-anchored horseradish peroxidase...mitochondria (Hoover et al., 2025). MitoTRACER relies on a Cre-lox switch to turn recipient cells from red to green...nuclease, and excises the DsRed-STOP to trigger expression of EGFP. Figure 2: Schematic of the MitoTRACER...Confocal image of CA2 neurons in a mouse brain slice expressing PinkyCaMP; scale bar 50 µm. Reproduced from Fink...
  19. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...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...
  20. 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...
Showing: 41 - 60 of 83 results