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We narrowed to 270 results for: Cre expression vector

Showing: 241 - 260 of 270 results

  1. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

    Type
    Blog Post
    Published
    Sept. 9, 2025, 1:15 p.m.
    ... promoter to drive expression of a fluorescent gene and packaged into viral vectors that can be administered...non-coding sequences of DNA that regulate gene expression by interacting with proteins such as transcription...enhancers are functional in every cell. Differential expression of transcription factors, which recognize and...mechanism that allows enhancers to regulate gene expression in a cell type-specific manner also makes them...tools for targeting those specific cell types to express exogenous genes — a feature the AIBS has capitalized...Genetic Tools Atlas. To evaluate the resulting expression of the fluorescent reporter, the brain tissue...Functional enhancer elements drive subclass-selective expression from mouse to primate neocortex. Cell Reports...

  2. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    Published
    Jan. 19, 2021, 2:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ...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...as 30% of all cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations...he or she can use to study it (as seen in the screenshot below). Currently, the page includes interactive... Tackling Cancers' Drug Resistance with a New Screening Kit Learn How the Clon Tracer Library Can Be Used...

  4. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...the phage display screens are subcloned (using FX cloning) into the expression vector pSb-Init, which attaches...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...to, like ones that are hidden in the molecular crevices proteins on the cell surface. Figure 1: ...threonines at positions that can be randomized for creating the libraries (Zimmermann et al., 2018). Find ...protein (sybody), ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated...the candidate sybodies in their coat protein to screen for binding to the target protein. The protocol...

  5. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...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... sets, you can increase confidence that the labeling observed represents the expression level of the target...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...

  6. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...that the percentage of dystrophin-expressing muscle cells increases over time, and all three groups confirm...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...confirm dystrophin expression via Western blot, at <10% of the wild-type level. Muscle histology is improved...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...

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

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ...marvel of DNA compaction, an enhancer can increase the expression of a gene that is millions of base pairs...genome math expression for NANOG enhancers, we’re ready for step 2: design a CRISPR screen to disrupt ...CRISPR screen. We start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an...Putting it all together, we get the following expression for step 1: (H3K4me1 + H3K27ac) – H3K4me3 The...stem cells, and conclude with a targeted CRISPR screen that can disrupt these enhancers. A real-world ...that have NANOG binding sites 2. Design a CRISPR screen to target and disrupt the NANOG enhancers 3. CRISPR...you’re interested in designing your own CRISPR screens for enhancers, check out the Appendix. Conclusion...

  8. 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...PAM site, Cpf1 cleaves in a staggered fashion, creating a 5 nucleotide 5’ overhang 18-23 bases away from...mediated editing, as Kleinstiver et al. saw no decrease in Cpf1 activity with 4-6 base deletions at the...

  9. 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 ...

  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...ve learned that increased accessibility is achieved through interconnected incremental changes. It was...When Addgene opened in 2004, we focused on creating a repository that made it easy to share plasmids,...identified and offered a number of solutions that increased accessibility, such as using couriers that refill...

  11. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...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...for quantitative imaging, Ming Hammond’s group created Spinach2 using site-directed mutagenesis of Spinach...for in vivo studies. Dimerization of Broccoli increases its fluorescence by about 1.8-fold, and may be...for greater contact with its RNA aptamer. This increased interaction between Corn and its small molecule... to TO1-biotin. Two of these variants exhibit increased brightness compared to Mango, and all are photostable...

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

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ... groups was to create a system that would use light to turn on and off gene expression while imparting...HEK293T cells to induce expression of human IL1RN, the researchers saw an 11-fold increase in mRNA levels after...Phillips-Cremins JE (2019) LADL: light-activated dynamic looping for endogenous gene expression control...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...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...

  13. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    Published
    Aug. 8, 2019, 1:19 p.m.
    ...changes in root-to-shoot ratio in Arabidopsis, (2) expression patterns in plants with enhanced sodium accumulation...protocols Find eBooks on plasmids, CRISPR, viral vectors, and more ...the community, increasing the impact of the work, and as a collateral benefit, increasing the impact of...Tompson Institute.  References Baldwin, Melinda. "Credibility, peer review, and Nature, 1945–1990." Notes and...

  14. 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...antibody” can be used for downstream IP/ChIP assays. Creating epitope-tagged clonal cell lines remains a challenge...crRNA complementary to the template strand could increase knock-in efficiency.Note: Our TAG-IN tool can ...

  15. 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...

  16. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    Published
    Dec. 12, 2019, 2:22 p.m.
    ...search scoped to plasmids shows filters for expression and vector type, while a search scoped to viral preps...well-characterized. During this time, we’ve also created a variety of educational resources in an effort...understanding of Addgene’s customer’s needs. Categories We created a categorization of our information in order to...for promoter and serotype, among others. We also created a normalization mechanism to “collapse” variations...

  17. 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...specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...knock in models). CRISPR mouse model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide...selection offered by mouse ESCs. For example, we often screen only 15-25 mice when generating knockout models...for generating mouse models Although CRISPR is incredibly useful for generating mutations by NHEJ and generating...

  18. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ...drawback of CRISPR-Cas9 technology is that most expression systems are constantly “on”. There is no mechanism....org  View Addgene's collection of empty gRNA vectors View Addgene's CRISPR resources Request ready-to-use... or phosphate molecules of the RNA backbone to increase stability.  Nature has evolved ways to prevent...surprisingly well-tolerated by the Cas9 protein and can increase on-target efficiency. In the same vein, locked...

  19. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    Published
    Dec. 6, 2022, 2:15 p.m.
    ...viral vectors (here and here).  Figure 3: JEDI-2P was identified from a high-throughput screen, selected...Figure 2: iGluSnFR3 displays enhanced brightness, expression, and sensitivity compared with earlier SF-iGluSnFR.... By fusing ITAM to an RTK of interest and co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence... They use a unique and powerful high-throughput screen to select variants based on performance metrics...resulting JEDI-2P has faster response kinetics and increased brightness, sensitivity, and photostability under...selected for increased voltage response, brightness, and photostability. It enables sustained deep-tissue... be generalized to other RTKs of interest, they create sensors for FGFR1, EGFR and ErbB2. And orthogonal...

  20. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...Edit Collection The CRISPR Guide Empty gRNA Expression Vectors... we performed high-throughput ‘self-targeting’ screens with oligo libraries where pegRNAs are coupled ...cell contexts. Figure 1: High-throughput screening of pegRNA efficiency and building prediction tools...sequence-based prediction models from self-targeting screens such as PRIDICT/PRIDICT2.0 provide valuable insights...your target. Figure 3: High-throughput screening of the effect of chromatin on prime editing efficiency...
Showing: 241 - 260 of 270 results