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We narrowed to 303 results for: crispr cas9

Showing: 61 - 80 of 303 results

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

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    Blog Post
    ...Institute's Silvana Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation... the CRISPR system was a large one.  The question for her then was this: Can you engineer CRISPR-Cas9 ...describe just such a system, which they call CRISPR/Cas9 Synergistic Activation Mediator or SAM. SAM is...Two-way CRISPR screening In October, Jonathan Weissman’s lab introduced additional pooled CRISPR libraries... transcriptional activation by an engineered CRISPR-Cas9 complex. Konermann S, Brigham MD, Trevino AE,...CRISPR technology has changed how scientists edit and control genes, but according to the Broad Institute's... three components: 1. A nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA incorporating two MS2 RNA...

  2. Sequencing Options for CRISPR Genotyping

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    Blog Post
    ...Use Cas9 Activators Perform Genome-Wide Screens with CRISPR Generate Mouse Models with CRISPR/Cas9 Additional...View Our CRISPR Guide Pages Find CRISPR Plasmids for Your Experiments Get Ready-to-Use CRISPR Lentivirus...steps in the CRISPR experimental process is validating edits. Regardless of which CRISPR genome editing...discussed in CRISPR 101: Validating Your Genome Edit, there are a variety of options for CRISPR genotyping...techniques, please see “Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity” by Tycko et al. 2016...and improving the genome-wide specificities of CRISPR–Cas9 nucleases”
by Tsai and Joung 2016. Many thanks...-throughput screening strategy for detecting CRISPR-Cas9 induced mutations using next-generation sequencing...

  3. Degrading DNA with Cascade-Cas3

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    Blog Post
    ... blog CRISPR 101: Cas9 vs. The Other Cas(s) CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems...methods rely on Cas9 or a derivative of Cas9, but the ever-expanding repertoire of CRISPR has brought many..., these types of CRISPR systems work using a multi-Cas complex called Cascade (CRISPR-associated complex...mechanical work for editing. Class 1 CRISPR systems make up the majority of CRISPR-based immune systems in bacteria...by anti-CRISPR proteins or running into other proteins naturally bound to the DNA. Cas3 or Cas9? Large ...achieved using Cas9, but Cas3 may be a better option. To get large deletions using Cas9, two or more gRNAs... Engineering Using CRISPR-Transposons Additional resources on Addgene.org CRISPR guide Cascade-Cas3 ...

  4. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

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    Blog Post
    ...  New to CRISPR/Cas9 Technology? Learn More: Addgene's CRISPR/Cas9 Guide History of CRISPR Cas - A tale...awards that will be bestowed upon discovers of CRISPR/Cas9 genome engineering, Dr. Jennifer Doudna and Dr... They are finding they have been able to use CRISPR/Cas9 to generate mice with genes knocked out in the...He offered the great analogy of comparing the CRISPR/Cas9 system to Norton Antivirus. Antivirus programs... crispr/cas9...Addgene Scientist Matt Ferenc (one of our resident CRISPR experts) at the 2014 Johnson & Johnson’s Dr. Paul...technologies. Addgene is proud to be helping get CRISPR reagents out to academic labs. It is our hope that...

  5. CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

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    Blog Post
    ...Addgene.org Check out CRISPR Topic Page Find CRISPR Nickase Plasmids Browse the CRISPR Guide ...rules for your next CRISPR nickase experiment Overview of Cas9 nickase We’ll use SpCas9 nickases as examples...By mutating one of two Cas9 nuclease domains, researchers created the CRISPR nickase. Nickases create...strands bluntly with WT Cas9 and one gRNA, you can create a staggered cut using a Cas9 nickase and two gRNAs...Integrated DNA Technologies) first demonstrated how CRISPR nickases improve homology directed repair rates...targeting range: using an individual gRNA with WT Cas9, repair levels decrease rapidly 10 bp from the cut...The HDR efficiency compared favorably to using WT Cas9 with the left gRNA across the window. In a subsequent...

  6. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

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    Blog Post
    .... More CRISPR Refereces: Browse CRISPR Plasmids CRISPR Information at Addgene More CRISPR-Cas Blog...of the CRISPR article "Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9" from the...Orkin and Daniel Bauer's labs details the use of CRISPR/Cas9 to create genomic deletions in mammalian cell...the motivations behind this research.   Using CRISPR/Cas9 for targeted genomic deletions We were inspired...with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore the paired double-strand...clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 nuclease system in mammalian cells. J.Biol.Chem...bear an indel or inversion mutation [3]. In the CRISPR era, the genomic landscape may be surveyed in unprecedented...

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

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    Blog Post
    ...Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease...Addgene.org Browse CRISPR plasmids Check out our CRISPR guide pages Find ready-to-use Cas9 lentivirus ...target site. With the widespread adoption of CRISPR/Cas9 technology and rapid improvements in the field...tracrRNA/Cas9 Ribonucleoprotein (csRNP) delivery Epitope tagging of mammalian genes using CRISPR depends... Figure 2: Cas9 RNP complexed with 2-part cr:tracrRNA tags genes more effectively than Cas9 complexed ...csRNP knock-in experiments If your lab is new to CRISPR/Cas9 experiments, there is no need to worry at all...Edinburgh. His research interests include optimizing CRISPR/Cas9 genome-editing methods and applying them to study...

  8. Tips for CRISPR Gene Editing in Mice

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    Blog Post
    ... encoding Cas9 into the oocyte. This original method was developed as the use of CRISPR/Cas9 in mammalian... by guest blogger Samantha Young. The use of CRISPR/Cas9 for gene editing has expanded since its adaptation...al., 2014). There are several variants of the CRISPR/Cas9 system floating around, and many pre-designed...running as soon as possible! Selecting your target CRISPR/Cas9 is often used to study the biology surrounding...Plasmids for Testing Your gRNAs in vitro at Addgene CRISPR/Cas9 gene editing in mice Now that you’ve validated...embryos. When researchers first started using CRISPR/Cas9 for genome editing in mice, they would often ...Design You gRNA Find CRISPR Plasmids for Use in Mammalian Cells Check out All of Our CRISPR Resources  ...

  9. X-CHIME: Context Dependent Germline Knockout in Immune Cells

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    Blog Post
    ...Editing CRISPR Cheat Sheat Resources from Addgene.org CRISPR Guide CRISPR: Protocol for Genome Deletions in... CHIME works through transduction of Cas9-expressing (Rosa26-Cas9) hematopoietic stem cells with a lentiviral...., Haining, W. N., & Sharpe, A. H. (2019). A CRISPR-Cas9 delivery system for in vivo screening of genes... Addgene blog CRISPR101: Validating Your Genome Edit How to Design Your gRNA for CRISPR Genome Editing...time-consuming, as it only changes one gene at a time. CRISPR-based systems enable gene knockout in immune cells...and in combination with different genes. Having CRISPR systems for assessing genes in defined contexts...genes pXPR_219 Violet-excited GFP (vex) Rosa26-Cas9 I-CHIME Inducible knockout of a single gene...

  10. Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring

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    Blog Post
    ....org Read our CRISPR guide Find CRISPR plasmids for your research Find links to CRISPR references ...hyperthermostable Cas9 CRISPR nuclease that is active up to 100°C.  The IgnaviCas9 protein is 1,241 amino acids...proteins Read all of our CRISPR blog posts Read about GeoCas9, another thermostable Cas9 Resources on Addgene.org...indicates that like GeoCas9 and ThermoCas9, IgnaviCas9 is a type II-C Cas9. IgnaviCas9 is most similar to...thermostable Cas9 enzymes have previously been characterized: GeoCas9 from the Doudna lab and ThermoCas9 from ...Find IgnaviCas9 here! What can you do with IgnaviCas9? Why would a thermostable Cas9 be useful? It can enable...launched in the Quake Lab. Identifying IgnaviCas9, a hyperthermostable Cas9 This year it was wonderful to learn...

  11. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

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    Blog Post
    ...Addgene.org Check out our CRISPR Guide Find Plasmids for Cas9 Purification Find CRISPR Plasmids for Mammalian...how you can make CRISPR mouse model generation easi-er! Generating mice before CRISPR The process of generating... process takes ~8-10 months. With CRISPR (B), you can inject CRISPR components directly into a zygote ...template and CRISPR machinery, Quadros et al. recommend ribonucleoprotein (RNP) complexes of Cas9 protein ...crRNA and tracrRNA) for maximum Easi-CRISPR efficiency. Easi-CRISPR is compatible with standard microinjection... 47327 pET-28b-Cas9-His 6xHis (C terminal); NLS (C terminal) 62933 pET-Cas9-NLS-6xHis 6xHis... 62934 pET-NLS-Cas9-6xHis 6xHis (C terminal); NLS (N terminal)   Easi-CRISPR efficiency and delivery...

  12. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast and then selected with... Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature...formed by expansion of single cells for 13 days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid...

  13. Plasmids for Endogenous Gene Tagging in Human Cells

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    Blog Post
    ...interest. To get the molecular components of CRISPR/Cas9 (Cas9 protein and the crRNA-tracrRNA complex) and...regulatory control. But even with the advent of CRISPR/Cas9 technology, inserting large tags into a precise.... We’ve created and tested plasmids that use CRISPR/Cas9 to endogenously tag a wide variety of genes with...HDR) process following the double strand break CRISPR/Cas9 makes at the target site. The whole segment—a... the plasmid from being cut and destroyed by CRISPR/Cas9 during transfection. Making the double strand...introduce the fluorescent tag to the cells, we used CRISPR/Cas9 to make a precise cut in the genome near either...Brock, et al. "Systematic gene tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization...

  14. 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...Limitation for CRISPR Delivery Genome-wide Screening Using CRISPR Performing In Vivo CRISPR Screens Using...development and improvement of CRISPR-based technologies over the past decade. CRISPR screens are easily customizable...integrate at double-stranded break points caused by CRISPR-Cas9 editing. The good news is that this integration...resources at addgene.org CRISPR pooled libraries Guide to pooled libraries CRISPR guide AAV guide ... interference, activation, and knockout. Most CRISPR screens rely on lentiviral systems, a fact supported...supported by the wide array of lentiviral-based CRISPR libraries available at Addgene. The large packaging...

  15. Top Requested Lentivirus and AAV of 2016

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    Blog Post
    ... provide a few non-CRISPR related viral preps. Cas9 is the headliner of most CRISPR applications and can...express Cas9 and a gRNA from the same plasmid. LentiCas9-blast virus can be used to generate Cas9-expressing...While various Cas9 mutants have been engineered to perform unique editing tasks, classic Cas9 nuclease, which...gRNAs in a monoclonal Cas9 background because you’ll have more consistent levels of Cas9 expression between...this plasmid Cas9 expression is typically cytotoxic, which makes generation of high titer Cas9 lentiviruses... expressed from lentiCas9-Blast. Purpose of LentiCas9-blast LentiCas9-blast is a Cas9 expression vector...Top Lentivirus: lentiCas9-Blast Most of Addgene’s lentiviral inventory is based on CRISPR tools, though ...

  16. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

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    Blog Post
    ...and efficiently, they’ve recently adapted the CRISPR/Cas9 system for protein tagging. We spoke with Mendenhall...go to www.addgene.org/crispr/tagging. Addgene: What motivated you to adapt CRISPR for use in protein tagging...context? Mendenhall: The benefits are two-fold: CRISPR/Cas9 offers high efficiency of inducing double strand...technologies. Generally speaking, anyone using CRISPR/Cas9 in the lab already should be able to run this...tagging with CRISPR Additional CRISPR Posts from the Addgene Blog ... crispr/cas9...Mendenhall and Myers about their new CRISPR tagging system, which they’ve now made available at Addgene...

  17. 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...blog Download the CRISPR 101 eBook Find our CRISPR 101 blog posts Read our CRISPR cheat sheet Resources...our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids Topics: CRISPR, Other...edition of “What’s New in CRISPR” - where we highlight a few of the newest CRISPR plasmids available at Addgene... read more content related to CRISPR technology, subscribe to our CRISPR blog content.   In this post ... Miniature CRISPR-Cas systems SEND plasmids Prime editing improvements Miniature CRISPR-Cas systems ...One drawback to the commonly used S. pyogenes CRISPR/Cas9 system is that it is too large to be effectively...

  18. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

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    Blog Post
    ...other CRISPR reporters Browse our CRISPR blog posts Resources on Addgene.org Browse the CRISPR guide... genome, but they’re inefficient compared to CRISPR/Cas9 edits that rely on double strand DNA breaks. ...single base editing by APOBEC-Cas9 Though the ACE reporter quantifies APOBEC-Cas9 efficiency in real-time, ...converted into BFP (blue fluorescent protein) upon CRISPR/Cas9 homology directed repair (HDR) (Glasser et al... A Versatile Assay for the Quantification of CRISPR/Cas9-mediated Genome Editing. Molecular Therapy - ...and enrichment of DNA editing by APOBEC–Cas9 or cleavage by Cas9 in living cells. Nucleic Acids Research... a APOBEC-Cas9n-UGI complex. So how exactly does this work? Figure 1: APOBEC- and Cas9-mediated ...

  19. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

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    Blog Post
    ...Most popular methods for CRISPR activation CRISPR activation uses dCas9, a CRISPR protein variant lacking... blog Browse our many CRISPR blog posts Get the basics of CRISPR in the CRISPR 101 blog series Resources...making it a common choice for CRISPR activation. Newer methods for CRISPR activation While SAM, SunTag,...compared to other CRISPR activators, but their initial results show promise for CRISPR-based gene activation... transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org...Gersbach CA (2013) RNA-guided gene activation by CRISPR-Cas9–based transcription factors. Nat Methods 10:973...Resources on Addene.org Read Addgene's CRISPR Guide Find CRISPR plasmids for your research ...

  20. CRISPR 101: Epigenetics and Editing the Epigenome

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    Blog Post
    ...types. They have also fused LSD1 to x-Cas9(3.7). x-Cas9(3.7) is a SpCas9 mutant that exhibits increased PAM...deacetylases. CRISPR and epigenetics As with many TALEN-based technologies, the advent of CRISPR has made .... For an up-to-date list of CRISPR epigenetic tools, check out our CRISPR Epigenetics resource. Transcriptional...potential advantage of CRISPR epigenetic tools is their persistence and inheritance. CRISPR activators and repressors...TE, Gersbach CA (2015) Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from ...IB, Crawford GE, Reddy TE, Gersbach CA (2017) CRISPR–Cas9 epigenome editing enables high-throughput screening...Edwards JR, Challen GA (2016) Reprogrammable CRISPR/Cas9-based system for inducing site-specific DNA methylation...
Showing: 61 - 80 of 303 results