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We narrowed to 224 results for: cas9 expression plasmid

Showing: 61 - 80 of 224 results

  1. What's New in CRISPR - September 2019

    Type
    Blog Post
    ...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...They’ve deposited plasmids for active and inactive CasX for E. coli and human expression. Read the paper...inducible CRISPR/Cas9 systems for transcriptional activation and genome editing by fusing Cas9 to a mutated...Nucleic Acids Research Find the drug inducible Cas9 plasmids E. coli genome-wide inhibition library In the...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...vectors to create knock-ins CasX Drug inducible CRISPR/Cas9 activation E. coli genome-wide inhibition library...vectors GeneWeld from Jeffrey Essner’s lab uses CRISPR/Cas9 and a donor vector to create knock-ins of fluorescent...

  2. What's New in CRISPR - November 2021

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    Blog Post
    ... enzyme with a fully active Cas9 enzyme. PE-Cas9 comprises an active Cas9 fused to the reverse transcriptase...restoring expression in the liver.  Read the paper in Nature Biotechnology Find the PEDAR plasmids   Additional...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids Topics: CRISPR, Other CRISPR... where we highlight a few of the newest CRISPR plasmids available at Addgene. If you want to read more...will cover: Miniature CRISPR-Cas systems SEND plasmids Prime editing improvements Miniature CRISPR-Cas...drawback to the commonly used S. pyogenes CRISPR/Cas9 system is that it is too large to be effectively...CasMINI system is less than half the size of the Cas9 or Cas12a based systems, making it well suited for...

  3. Addgene's Top 10 Blog Posts of 2015

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    Blog Post
    ...control gene expression. Get the full scoop from Addgenie A. Max Juchheim in this post. 3. Plasmids 101: E.... the strains you can use to express your protein of interest. 4. Plasmids 101: Blue-White Screening Getting...While the vast majority of CRISPR plasmids in our repository use Cas9 to cut DNA, Addgene Science Communications...CRISPR mediated genome engineering. 10. Plasmids 101: Control Plasmids Need help thinking about the types ...Scientists eBook Download the Plasmids 101 3rd Ed. eBook Read the Most Recent Plasmids 101, CRISPR 101, and Career... two eBooks (Management for Scientists and the Plasmids 101 2nd ed, the 3rd edition has now been released...top of the many developments in science and the plasmids, technologies, and researchers behind them. As...

  4. Using Addgene's New Viral Service for Your Research

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    Blog Post
    ...preps for the delivery of Cas9 endonuclease, Cas9 nickase, Cas9 activators, Cas9 repressors, select gRNAs...accelerate your research.   Why distribute virus? Plasmid distribution is booming, but, over the years, many...list of what’s available or suggest additional plasmids with the Viral Service Suggestion Form. Lentivirus... to knockout a gene, replace it, modulate its expression, or perform a genome-wide screen, we’ve got viruses...experiments, while rtTA can be used to activate expression of a gene under the control of the tet operator...page and includes western blots as well as other expression data. If you have any questions about the Viral...Uses and Overview AAV: A Versatile Tool for Gene Expression in Mammals Additional Resources on Addgene.org...

  5. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

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    Blog Post
    ...combining specific sgRNAs with Cas9 or Cas9 derivatives. Unmodified Cas9 causes double stranded DNA breaks... loss of function while nuclease-deactivated Cas9 (dCas9) can be tethered to a repressor (CRISPRi) or ...sgRNA library In addition to inhibiting gene expression, dCas9 can also be used for transcriptional activation... activation domain (CRISPRa) to regulate gene expression. Some CRISPR libraries contain many sgRNAs per... issues. CRISPRi libraries utilize an engineered Cas9, which contains multiple point mutations in the ...screens can help reveal the function of lowly expressed genes or genes that are more drastically affected...) . This tracrRNA was combined with the sgRNA expression vector allowing the screen to be conducted with...

  6. Degrading DNA with Cascade-Cas3

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    Blog Post
    ...deletions. Many of these methods rely on Cas9 or a derivative of Cas9, but the ever-expanding repertoire of...bound to the DNA. Cas3 or Cas9? Large genomic deletions can be achieved using Cas9, but Cas3 may be a better...previously mentioned issues with expressing all Cascade components can make Cas9 easier to work with than Cas3...its partner in crime, Cascade. Find Cascade-Cas3 plasmids in Addgene's collection! A “complex” system Cas3...challenging. It can be difficult to ensure comparable expression of all components of Cascade for maximum efficiency... Various labs have designed all-in-one Cascade plasmids — such as the Tomoji Mashimo lab and the Tilmann...efficiency of these systems, though consistent expression can still be a challenge. A multitasking Cas ...

  7. Open Resources and Plasmid Tools For Studying C. elegans

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    Blog Post
    ... a GAL-4 based bipartite expression system that can regulate gene expression in C.elegans. In this system... cGAL can drive expression of your gene of interest.  SapTrap Tool Kit for CRISPR Cas9 edits At Addgene... CRISPR plasmid can be created in a single tube. You can also find additional SapTrap plasmids used for...about C. elegans and the plasmids we have at Addgene check out our worm expression resources page. This page...about SapTrap Read about CRISPR/Cas9 in C. elegans Browse our Plasmids 101 blog posts Resources on Addgene.org...CRISPR techniques and new tools for conditional expression and degradation. These workshops not only highlighted...discussed at the meeting, in addition to useful plasmids and toolkits available at Addgene.  C. elegans...

  8. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    ... smaller size of the plasmids. The Zhang lab, on the other hand, uses a 1 plasmid, single infection system...have previously been stably transfected for Cas9 expression. The Yusa lab has used their library to screen...It’s clear that CRISPR-Cas9 technology has really changed the game for anyone looking to quickly and ... thousands of targets at once. That’s what our plasmids allow.” Wang’s library is designed to target a...Lander library uses sequential infection first of Cas9, then with the sgRNAs, and can result in a higher...application offer proof-of-principle that CRISPR-Cas9 can be used to target any sequence in the genome... Genetic Screens in Human Cells Using the CRISPR/Cas9 System. Wang T, Wei JJ, Sabatini DM, Lander ES. ...

  9. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ... to use variants of Cas9, like split Cas9, or a smaller Cas9 ortholog like SaCas9. You could even venture...to use a multi-plasmid system, where you have one AAV vector dedicated to expressing Cas9, while the other...Addgene. The large packaging capacity and stable expression of lentiviral vectors make them an attractive...attractive option. However, safety requirements and expression limitations may be too constraining for some experiments...improved safety profile, customization, and enhanced expression in many mouse tissues. Safety AAV and lentivirus...significant degree using different types of AAV. Expression Another advantage of AAV vectors is how the cargo...cargo — in this case, the CRISPR components — are expressed. Unlike lentivirus, cargo delivered by AAV does...

  10. Hot Plasmids February 2024

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    Blog Post
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...facilitates endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in ...all applications. Addgene’s plasmid collection includes over 10,000 plasmids with a FLAG tag (!), but did...lines and primary cells. Although cell entry of a Cas9 ribonucleoprotein (RNP) was limited (likely due ...multiplexed guides. Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization...Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral sgRNA targeting mCherry reporter... al. 2023 with permission.   Find Cas-PAGE plasmids here!   Zhang, Z., et al. (2023). Efficient engineering...

  11. Hot Plasmids: Spring 2025

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    Blog Post
    ...empty and EGFP retroviral plasmid set, our bacterial plasmid set for expressing tagged proteins, and stay...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. ...backbones special: Strong expression: A CMV promoter drives robust gene expression in mammalian cells. Flexible...stay tuned for mammalian expression and AAV plasmid sets launching later this year!  Find Addgene's own...) Fluorescence from axonal boutons of neurons expressing iGluSnFR3, iGluSnFR4f, and iGluSnFR4s. C) Spike-triggered... synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A., Negrean, A., Chen, Y., Iyer...powerful tools in a scientist's toolkit is the empty plasmid backbone: the DNA vector that contains everything...

  12. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

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    Blog Post
    ...to edit a gene, the expression of Cas9 is no longer necessary. However, Cas9 can be toxic if continuously...leading to transient expression of gene products (Wu et al., 2004). Over time, the expression diminishes as ...2008). 2. Transient expression of transgenes/shRNAs If you want your cells to express a protein or nucleic...the expression of the delivered payload is gradually reduced. For example, after using CRISPR/Cas9 to ...continuously expressed (Fu et al., 2013). Using IDLVs ensures transient expression without the risks associated...associated with prolonged expression. 3. Stable expression in non-dividing cells IDLVs can also be useful for...integrase-deficient packaging plasmid instead of the standard packaging plasmid. For more details on how to...

  13. Mapping the 4D nucleome with CRISPR/Cas9

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    Blog Post
    ... a colorful Cas9 toolbox, Ma et al. turned to SpCas9 and its orthologs NmCas9 and St1Cas9. Each ortholog...states. dCas9 gets a colorful makeover Even without its nuclease activity, catalytically dead Cas9 (dCas9... colorful system that uses fluorescently labeled Cas9 to label multiple genomic loci in live cells. While...hybridization (FISH) or fluorescently labeled TALEs, CRISPR/Cas9’s ease of use and ability to label live cells make... chromatin organization. This fluorescent CRISPR/Cas9-based method is a great step forward in live cell...As this study joins the pantheon of useful CRISPR/Cas9 techniques, we at Addgene are excited to see what....1420024112. Epub 2015 Feb 23. Pubmed. Find the plasmids from this publication at Addgene. Dynamic imaging...

  14. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

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    Blog Post
    ...Silvana Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation in mind...tetraloop and stem-loop 2; 3. The MS2-P65-HSF1 plasmid which expresses the activation helper protein. SAM can ...question for her then was this: Can you engineer CRISPR-Cas9 activators that work well enough on any gene that...describe just such a system, which they call CRISPR/Cas9 Synergistic Activation Mediator or SAM. SAM is an... three components: 1. A nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA incorporating two MS2 RNA...system, Konermann explained, was new knowledge about Cas9’s crystal structure. “I thought maybe we could use...levels of control in the system. First of all, the Cas9s can be turned on and off, to reversibly activate...

  15. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    ...Postdoctoral Researcher in Jacob Corn’s lab. CRISPR-Cas9 (Cas9) is an RNA-guided nuclease that targets and cuts...replacement by HDR. Understanding Cas9 biophysics enhances HDR We first measured Cas9 dissociation rates and discovered...of reagents is relatively modest, requiring expressed Cas9 protein, transcribed sgRNA, and optional donor...interaction with the Cas9-DNA duplex. Image from Richardson et al., 2016.   dCas9 enables sequence replacement...normally accompanies Cas9 cutting. We still don’t know the mechanism underlying dCas9 editing, but it could... cuts genomic DNA. The interplay between Cas9 (which causes the breaks) and host cell DNA repair factors...factors (which repair those breaks) makes Cas9 extremely effective as a genome editing reagent. This interplay...

  16. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...catalytically active Cas9, the short, CRISPRainbow gRNA will permit labeling, but a newly expressed standard length...PMC4864854. Find the plasmids from this publication at Addgene. 4. Kiani, Samira, et al. “Cas9 gRNA engineering... a colorful Cas9 toolbox, Ma et al. turned to SpCas9 and its orthologs NmCas9 and St1Cas9. Each ortholog...locus, and to do so in vivo. Catalytically dead Cas9 (dCas9) cannot induce gene editing, but it retains its... All 6 gRNAs can be expressed in a single vector, pCRISPRainbow-DONOR1, with dCas9 supplied by a separate...success of Ma et al.’s approach, fluorescently labeled Cas9’s come with a number of limitations. Since color...by PAM sequence, each color requires a different Cas9 ortholog, and the target sequence must be located...

  17. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

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    Blog Post
    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...about a recent plasmid deposit please sign up here.   Listen this episode of Hot Plasmids! Light controls...available in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). Fabritius...APOBEC- and Cas9-mediated Editing (ACE) in real-time. The reporter constitutively expresses eGFP to indicate...Hahn's lab at the University of North Carolina (plasmids at Addgene). It consists of two domains, LOV2 ... the same number of fluorophores. You can find plasmids containing the cobalamin binding riboswitches ...

  18. Some Like it Hot: Thermostable GeoCas9

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    Blog Post
    ... 1. Comparing SpCas9 and GeoCas9. *For in vitro Cas9 RNP activity or longevity. GeoCas9 is active at high...used to make eSpCas9 and SpCas9-HF1. By mining the treasure trove of naturally occurring Cas9 diversity ...Activity is expressed as Kcleave (min-1). SpCas9 has maximum activity from 35-45 °C, while GeoCas9 has maximum...bacteria, such as Streptococcus pyogene’s SpCas9. These Cas9 proteins don’t work well at high temperatures...discovery of GeoCas9 by the Doudna lab has opened up the field of thermophilic bacteria to CRISPR/Cas9 genome...discovered a Cas9 protein from the thermophilic bacteria Geobacillus stearothermophilus (GeoCas9). G. stearothermophilus...human plasma. The discovery of GeoCas9 clears the way for CRISPR/Cas9 genome editing in facultative and...

  19. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...needed by Cas9 for cleavage. In certain cases, Cpf1 may be better suited for genome editing than Cas9 - read...CRISPR system. Like Cas9, Cpf1 contains a RuvC-like endonuclease domain, but it lacks Cas9’s other HNH endonuclease...Potential advantages of Cpf1 over Cas9 Type II CRISPR systems based on Cas9 were thought to be the simplest...than that of the widely used SpCas9 and comparable to SaCas9. As seen with Cas9 orthologs, Cpf1 efficiency...rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9-HF1. Both AsCpf1 and LbCpf1 ribonucleoproteins...needed for Cas9 function. Since both Cpf1 and its guide RNAs are smaller than their SpCas9 counterparts...display cleavage activity in mammalian cells. Like Cas9 nucleases, Cpf1 family members contain a RuvC-like...

  20. The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years

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    Blog Post
    ...with all the new plasmids. So far, we have distributed over 10,000 CRISPR/Cas9 plasmids to more than 3,...for shRNA expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're...packaging and expression vectors, including pMD2.G and psPAX2. Actually, these two plasmids from Didier ...influx of plasmids for reprogramming differentiated cells into pluripotent stem cells. Plasmids from Shinya... have 19 different ubiquitin plasmids in the repository. 100,000 Plasmids Shipped! - By 2010, Addgene ...our stride. We shipped our 100,000th plasmid. A variety of plasmids continued to be popular years after...as pBABE-puro for retroviral expression and pLVTHM, a lentiviral plasmid for mammalian gene silencing....
Showing: 61 - 80 of 224 results