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We narrowed to 294 results for: expression vector

Showing: 261 - 280 of 294 results

  1. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    Published
    June 22, 2023, 1:15 p.m.
    ... on the Addgene blog Viral Vectors 101: Types of Viruses Viral Vector Elements Tips for Preventing... contact with than GFP. Similarly, if the viral vector carries an shRNA to a gene essential for cellular...cellular function, this would also be a higher risk vector. While it is unlikely that you, or anyone else ...being integrated by the virus. If an oncogene overexpression construct is being delivered, that is a higher...

  2. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...myoblasts; when injected into DMD mice, these cells expressed functional dystrophin. Editing genes in vivo is..., the CRISPR components would only need to be expressed for a short period of time, which would prevent...et al. show that the percentage of dystrophin-expressing muscle cells increases over time, and all three...three groups confirm dystrophin expression via Western blot, at <10% of the wild-type level. Muscle histology...stem cells 2. Remove any immunogenicity of the AAV vector Once delivery has been optimized, it’s important... reduce off-target editing. Short-term CRISPR expression would be especially desirable, as it would reduce... genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy. Science...

  3. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...deliver in low-capacity vectors, such as adeno-associated viral (AAV) vectors. In 2017, Zetsche et al....multiplexing approach using a single crRNA array to express up to 4 crRNAs. Zetsche et al. also suggest that...

  4. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    Published
    Sept. 28, 2023, 1:15 p.m.
    ... of katanin to that anchor. So, you’ll need to express both the EB3N-VVDfast-iLID anchor module and the...specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID along with your...Opto-RhoGEF design and activity. B) OptoTIAM (magenta) expressed in a monolayer of endothelial cells, showing cell...either CRY2 or CIB1 to your protein of interest and express the other separately, then use blue light to generate...probe GFP-PASS. B) Time series of HEK 293T cells expressing plasma membrane-targeted SuperPLDx30 (magenta...Additional resources on the Addgene blog Viral Vectors 101: Optogenetics Additional resources on Addgene.org...

  5. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ... DNA compaction, an enhancer can increase the expression of a gene that is millions of base pairs away...Putting it all together, we get the following expression for step 1: (H3K4me1 + H3K27ac) – H3K4me3 The...enhancers with CRISPR Armed with the genome math expression for NANOG enhancers, we’re ready for step 2: ...of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated retrovirus in the same family as...

  6. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...al., 2024). In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study...than SpyCas9, allowing them to be packaged in AAV vectors, potentially with multiplexed targets.   ...showed that AAV-delivered CHARM could silence the expression of prion protein in mouse brains for at least...gene product; it simply turns the target protein expression off. Plus, DNA methylation is a long-lasting ...-silencing CHARM variants that stop their own expression after silencing their target — potentially improving...by targeting marker proteins that are uniquely expressed in a given cell type. For example, glial fibrillary...GFAP) is a glia-specific intermediate filament expressed in astrocytes throughout the brain, making it ...

  7. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ... of interest. In contrast, RNA guides must be expressed from a plasmid or in vitro transcribed, and changes...advantageous if you’re working with small capacity viral vectors such as AAV. Base removal RNA-guided CRISPR enzymes...

  8. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...step as simple as heat-shocking to induce the expression of a Cre recombinase contained within the knock-in...modular toolkits build single plasmid targeting vectors that encode the gRNA transcript and the repair ...using modified Cas9 proteins to regulate gene expression (CRISPRi/CRISPRa), visualize specific genomic....114.170423 Schwartz ML, Jorgensen EM (2018) SapTrap vectors for introducing point mutations with unc-119+ selection...

  9. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ... library via a lentiviral vector to cell lines that either stably expressed Cas9 or that had been lentiviral...control for many of the effects of gRNA and Cas9 expression, they fail to account for the effects of Cas9...sites should control for the effects of guide expression and dsDNA breaks. Figure 2: Non-targeting...

  10. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ... lentiviral packaging systems, and luciferase expression. On individual material pages, we added curation...curation fields for purpose, promoters, expression types, and more, all of which help a researcher understand... Inducible Plasmids Empty Backbones Bacterial Expression Systems AAV Plasmids Blogs Origin of ... CRISPRTwenty Years of Sharing: Addgene's Viral Vector Service...

  11. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...SELEX, the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches can...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...

  12. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ... that would use light to turn on and off gene expression while imparting spatiotemporal control, reversibility...transcriptional activator domain (VP64 or p65). Upon expression in the cell, the dCas9-CIB1 fusion binds to the...exposure (470nm), the researchers showed that expression of the human ASCL1 gene could be spatially controlled... Using this system in HEK293T cells to induce expression of human IL1RN, the researchers saw an 11-fold...demonstrated the ability to spatially control gene expression. Both of these techniques take advantage of light-induced...light-activated split-Cas9 (FAST) system constitutively expresses the C-terminal fragment of Cas9 while transcription...N-terminus is light-dependent (Yu et al., 2020). To express the N-Cas9, Haifeng Ye’s lab used the bacterial...

  13. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...and Cas9-expression plasmids. Cas9 guide RNA delivery does however rely on cloning into a vector or in vitro...co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression plasmid to edit a gene in vivo. Try NgAgo for...

  15. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ...loxP sites such that their expression is turned on or off in cells expressing Cre. The result of a system...allows the visualization of fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell...immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph nodes...efforts have been made to create transgenic mice expressing fluorescent protein reporter constructs in the...above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after...Subcellular Localization Find Fluorescent Protein Empty Vectors ...

  16. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...pitfalls: they require construction of plasmid vectors and/or rely on cumbersome in vitro synthesis of...purchased and there is no need to construct plasmid vectors or synthesize/purify IVT sgRNA in the lab. Figure... (although we could successfully knock-in non-expressed genes with similar efficiency, Dewari et al. 2018...tagging success, although many of these genes were expressed at really low levels so actual knock-in success...

  17. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...targeted repression of gene expression. They successfully knock down expression of essential and non essential...) technology (Ronda et al., 2016). pMA7CR_2.0 expresses lambda Red and Cas9, which are separately inducible...function in bacteria, most efforts to regulate gene expression were limited to inducible promoters or direct... much more user-friendly way to modulate gene expression. The Marraffini lab and Qi lab developed early...repression. Using a Tet responsive promoter to drive expression of a catalytically dead Cas9 enzyme allows reversible...the gRNA cassette, one can reliably knockdown expression of multiple target transcripts simultaneously...genes in M. tuberculosis. dCas9 and a gRNA are expressed on pRH2502 and pRH2521, respectively. Both dCas9...

  19. Reagent Repositories Are Speeding up Science During the Pandemic

    Type
    Blog Post
    Published
    Aug. 4, 2020, 1:15 p.m.
    ...-2 are the lentiviral production and packaging vectors VSV-G and psPAX2, which were used in a publication... human cells via hACE2, could not enter cells expressing murine ACE2 (Subbarao et al., 2006). Now that...

  20. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    Published
    July 12, 2016, 2:30 p.m.
    ...Gurumurty et al., 2016). AAV can also be used as a vector to deliver CRISPR/Cas9 either through injection...Mediated Genome Editing with Adeno-Associated Viral Vector. iScience 9:286–297 . https://doi.org/10.1016/j.isci...Resources on the Addgene Blog CRISPR 101: Mammalian Expression Systems and Delivery Methods Learn about Mouse...
Showing: 261 - 280 of 294 results