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We narrowed to 422 results for: expression vector

Showing: 261 - 280 of 422 results

  2. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...gene drive is also the easiest: don't use a DNA vector that encodes both Cas9 and sgRNA. What if I want...spread exclusively through populations that already express Cas9 from an unlinked locus, while a Cas9+sgRNA...attempting to build gene drives in tropical mosquito vectors of diseases such as malaria and dengue are currently...

  3. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Cells You may also like... AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference...transfection: pHelper; pRC (Rep-Cap), plasmid expressing your gene of interest Triton X-100 Benzonase/...

  4. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    ...Fujii: We generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral...plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene: Your ...retroviral enChIP system enables us to express the CRISPR complex much easier in various cells, including primary... IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. Fujita T, Fujii...

  5. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...stability. Additionally, the choice of vector can impact the expression level of an antibody (Ayyar et al....such as parasites or microbes, or to proteins expressed on the surface of cells that have been infected...the protein out of solution, or isolate cells expressing this protein. Read on to learn more about antibodies...context of the immune system, each isotype is expressed at different times of the immune response and ...qualitative and quantitative measurement of protein expression in cell lysates, whole cells, or tissue samples...frequently used to compare relative levels of protein expression between cell types or treatment conditions. They....    Figure 4: Western blot showing GFP expression in cells that have been transfected with a plasmid...

  6. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...modify the genome but still want to affect gene expression? dCas9 can be used as a platform to deliver various...activation domains to the same genetic locus via co-expression of an epitope tagged dCas9 and antibody-activator...experiment, select from a variety of empty sgRNA vectors and design your sgRNA targeting sequence using ...Validated gRNAs for Your Next CRISPR Experiment Cas9 expression systems Addgene carries a variety of Cas9 containing...containing plasmids that have been optimized for expression in different species and cell types including...systems can be found here “CRISPR 101: Mammalian Expression Systems and Delivery Methods”. Validating your...

  7. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...consist of a group of genes/proteins that, when expressed, normally encourage cell survival, growth and ...Clone Collection 2.0. The collection is a set of vectors for use with the Gateway Cloning Platform (Life... is that it enables the simple construction of vectors to study the function of any of these genes through...

  8. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...are subcloned (using FX cloning) into the expression vector pSb-Init, which attaches a Myc-tag and a His6...their small size, nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target...sybodies can be subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for the production of tag-free proteins...

  9. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...flow cytometry, and mRNA expression.   If your protein of interest is expressed at similar levels in ...useful to produce a sample with mixed expression, with the expression measurable through an epitope tag such...epitopes. Detection of overexpressed (tagged or untagged) protein When expression plasmids are available...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...caused by overexpression. Additionally, if the cell line used has endogenous levels of expression, this can...preferred. The overexpression approach is therefore best suited to cell lines without any expression of the target...antibody to detect the antigen of interest when expressed at endogenous levels (Laflamme et al., 2019). ...

  10. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ... 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...

  11. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...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...

  12. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...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...

  13. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ... 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...

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

    Type
    Blog Post
    ... 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...

  15. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...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 ...

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

    Type
    Blog Post
    ... 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...

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

    Type
    Blog Post
    ...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...

  18. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ... 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...

  20. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ... 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...
Showing: 261 - 280 of 422 results