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We narrowed to 263 results for: Cre expression vector

Showing: 241 - 260 of 263 results

  1. 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...

  2. 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 ...

  3. 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...

  4. 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...

  5. 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...

  6. 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...

  7. 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 ...

  8. 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...

  9. 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...

  10. 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...

  11. 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...

  12. 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...

  13. 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...

  14. The Challenges of Cell Culture

    Type
    Blog Post
    Published
    May 5, 2016, 2:30 p.m.
    ...along with Down-Regulation of HIF-1α by NDRG2 Expression Attenuates Proliferation and Invasion in Renal...on the Addgene Blog Learn All about Mammalian Vectors Read up on CRISPR Delivery Systems for Mammalian...line you will need to grow this initial stock and create a set of vials of ‘Master’ stock, which can be ...thawed, it is time to reorder this cell line to create new Master and Working stocks to work from. The...

  15. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 3, 2022, 1:15 p.m.
    ... vitro, and can be used for measuring protein expression or localization in cells, or even single-particle...Mol Syst Biol. 2014 Galit Lahav EYFP Gateway vectors with EYFP split at 175 Arabidopsis homolog of the...acetylation system and novel in vivo high-throughput screen for molecules affecting alpha-synuclein oligomerisation...

  16. Lucky Thirteen: Guest Contributors of 2023

    Type
    Blog Post
    Published
    Dec. 5, 2023, 2:15 p.m.
    ...Information DatabaseResearcher: Amos BairochViral Vectors 101: Systemic CapsidsResearchers: Cynthia Arokiaraj...celebrations! We're kicking it off by sharing all the incredible guest posts and stories featured in our blog ...Reporter Genes: Ultrasound Imaging of Gene ExpressionAuthors: Marjorie Buss and Mengtong "Tom" DuanVisualizing...Plasmid Request Data Author: Joachim Goedhart  Created in collaboration with Addgene writers High Complexity...

  17. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...manipulated by injected RNA and can express fluorescent proteins. This lab has created an automated tank for feeding...continues to be challenging. Organism specific viral vector systems would also be beneficial for many systems... progress on the NSF “G to P grand challenge.” Creating a productive, new scientific community is no small...in particular, that they all share the plasmids created under this programs via Addgene. New organisms ... organisms) -  perhaps you’ll choose to study Crepidula atrasolea or Prochlorococcus with your next project...crisis. I like how they are using protocols.io to create a protocol community! Asclepias syriaca and Asclepias...used to study aspects of cognition. Spiralians - Crepidula atrasolea (Jonathan Henry) This black footed slipper...

  18. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...with natural chain termination at stop codons, expression levels of these tagged proteins are often very...could be substantial: Plasmid tRNA Synthetase Expression Host pMAH-POLY Polyspecific aminoacyl-tRNA ...intact cells. The Gradia Lab has deposited a gateway vector with Addgene, where you can insert your protein... drug discovery by big pharma (i.e. HTRF and LanthaScreen) (Degorce et al., 2009), measuring intracellular...are critical in setting up a FRET-based assay or screen (the Addgene blog has an excellent primer on FRET... biomolecules each containing multiple binding crevices that can nonspecifically soak up your fluorophore...donors (Fig. 1). Scores of FP variants have been created, thereby enabling a broad spectrum of donor fluorescence...

  19. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ... whatever molecule you choose (Figure 1). Just express the fusion protein and add a specific ligand! A...better detect dim structures or low levels of expression with the same imaging workflows you already use...to drive cell type-specific delivery of a viral vector (Strebinger et al., 2023). All this flexibility...no wonder that self-labeling protein tags are increasingly popular. Embrace the flexibility and switch ...

  20. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    Published
    Feb. 8, 2024, 2:15 p.m.
    ...cleave DNA and instead activates/represses gene expression by various mechanisms. The RNA guide is typically...- GUIDE-Seq and integrase-defective lentiviral vectors - which rely on insertion of DNA by the NHEJ pathway...differences. If you are performing CRISPR library screens, the same principle holds true - the higher the...lower the likelihood of an off-target artifact. Increasing the N in your experiment is the only way to effectively...effectively control for off-targets and significantly decrease the probability of artifacts. Off-target controls...Rabinowitz R, Offen D. Single-Base Resolution: Increasing the Specificity of the CRISPR-Cas System in Gene...
Showing: 241 - 260 of 263 results