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We narrowed to 274 results for: crispr cas9 plasmids

Showing: 101 - 120 of 274 results

  1. PITChing MMEJ as an Alternative Route for Gene Editing

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    Blog Post
    ...Addgene.org Learn All about CRISPR on Our CRISPR Guide Pages Find CRISPR Plasmids for Your Experiments Deposit...This PITCh plasmic is cotransfected with a plasmid carrying Cas9, the PITCh-gRNA, and the locus-specifc gRNA...containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector carrying Cas9 and ...MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118...mammalian genomes as assessed by CRISPR design tools at crispr.mit.edu. They tested their system in HEK293...If you follow CRISPR research, you know all about using non-homologous end-joining (NHEJ) to make deletions...lab has harnessed MMEJ to create a new method for CRISPR gene knock-in, termed PITCh (Precise Integration...

  2. Targeted Mutagenesis with EvolvR

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    Blog Post
    ...nicking Cas9. A guide directs Cas9 to a site of interest, where it nicks its target DNA (left). Cas9 dissociates...Addgene.org Browse our CRISPR guide pages Find bacterial CRISPR plasmids ...within a user-defined genomic region. EvolvR, a CRISPR-Cas9 based targeted mutagenesis method developed by...specific mutations in your plasmids Use REPLACER Mutagenesis to mutate your plasmid Resources on Addgene....error prone PolI3M that’s fused to a nicking Cas9 (nCas9). Similar to mutagenesis with error prone PCR... targeted mutagenesis experiments? Find EvolvR plasmids at Addgene! References Gaj, Thomas, and Pablo...Pablo Perez-Pinera. "The continuously evolving CRISPR barcoding toolbox." Genome biology 19.1 (2018): 143...

  3. Addgene @ Keystone: Thoughts on Precision Genome Engineering and Synbio

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    Blog Post
    ...improving the CRISPR-Cas9 system. Mehmet Fatih Bolukbasi from Scot Wolfe's lab described a chimeric Cas9 that ...off our new CRISPR page (which describes how to plan your CRISPR experiment for our new CRISPR/Cas users...Find more CRISPR and Genome Engineering Resources at our website: CRISPR Resources & Plasmids Plan Your...followed by three of the biggest names in the CRISPR-Cas9 field – Jennifer Doudna, Feng Zhang, and Keith... Your CRISPR Experiment Synthetic Biology Resources & Plasmids  ...genome engineering from ZFNs through TALENs and CRISPR-Cas nucleases, provided important context for the...on this field. Advances in genome engineering & CRISPR technology That afternoon's workshop, entitled ...

  4. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...our comprehensive CRISPR guide Find CRISPR plasmids and resources Find CRISPRa plasmids ...blog Browse our CRISPR blog posts Download our CRISPR cheat sheet Download our CRISPR eBook Resources...What is CRISPR-based activation? To summarize, CRISPRa uses a nuclease-dead variant of Cas9 (dCas9) and ... for sensitive applications such as CRISPR screening. The plasmids used by the Qi Lab for this particular...recent papers describe the development of CRISPR activation (CRISPRa) tools for neuroscience, including transgenic... using CRISPRa to target individual genes can be relatively laborious because multiple plasmids/viruses...SPH mice CRISPRa has largely been limited to in vitro settings due to the large size of Cas9 activators...

  5. Another Pathway into Cells: iTOP

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    Blog Post
    ...iTOP would be compatible with CRISPR/Cas9 genome editing. Although CRISPR/Cas9 is all the rage, they had ...transduction method and its potential applications to CRISPR/Cas9 genome editing.   Protein-only transduction...expression. Image courtesy of Diego D'Astolfo. CRISPR/Cas9: a potential application of iTOP Having tested...stemness, indicating that iTOP may be valuable for CRISPR/Cas9 stem cell editing. The small molecule ...expression has a long-lasting effect, such as Cre- or Cas9-mediated recombination. Both primary hESCs and immortalized...cells treated with iTOP efficiently took up both Cas9 protein and a gRNA (yes, iTOP can also be used for... elements of iTOP. iTOP can be used to transduce Cas9 and a gRNA, which subsequently enter the nucleus...

  6. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

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    Blog Post
    ...using the CRISPR-Cas9 system CHIME Article contributed by Aliyah Weinstein  Listen to the CRISPR podcast...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... recent plasmid deposit please sign up here.      Listen to this episode of the Hot Plasmids podcast ... (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These ARG plasmids contain... custom nucleases for each gene of interest. CRISPR-Cas9 tools have been used in immunology for in vitro...Medical School developed a solution: CHIME, a CRISPR-Cas9 system that can edit genes in any immune lineage...

  7. 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...to use a multi-plasmid system, where you have one AAV vector dedicated to expressing Cas9, while the other...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...

  8. CRISPR-based Genome Editing Systems For Editing in E. coli

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    Blog Post
    ...Addgene blog: Browse CRISPR articles from our CRISPR topic overview Find other CRISPR methods for bacteria...Resources on Addgene.org: Visit the CRISPR Guide Get CRISPR plasmids from Addgene ...integration using CRISPR-associated transposases” or MUCICAT. Here, we’ll describe each of these CRISPR systems....pTargetF system consists of one plasmid that expresses the S. pyogenes Cas9, a temperature-sensitive replicon...beneficial for plasmid clearance. It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only.... pCutamp cures the plasmids by targeting the ampR promoter in all three plasmids. pCutamp is then removed... 13 plasmids with 14 rounds of transformation in 30 days. Figure 2: (A) MUCICAT plasmids for ...

  9. 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...Resources on Addene.org Read Addgene's CRISPR Guide Find CRISPR plasmids for your research ...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...

  10. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...Addgene.org Catch up on your CRISPR background with our CRISPR Guide Browse CRISPR plasmids for base editing Find...off-target editing based on Cas9 off-target activity.  Find CRISPR base editing plasmids at Addgene Improving..., a base editor containing high fidelity Cas9 variant HF-Cas9 (Rees et al., 2017). HF-BE3 showed 37-fold...other methods. In a head-to-head comparison with Cas9, Cas9 modified 9/12 known off-targets with a 14% indel...genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Science (New York, N.Y.), 368(6488)...individual Cas9. They also mutagenized the cytidine deaminase portion of the base editor to create SpCas9 base...et al., 2020 SpRY Cas9, available fused to ABEmax(7.10) or to BE4max   Evolved SpCas9 with NRN or NYN PAM...

  11. 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...

  12. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

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    Blog Post
    ...Addgene.org: Find RNA Knockdown Plasmids Check out our CRISPR Guide Read our CRISPR History  ...Microbial CRISPR diversity is impressive, and researchers are just beginning to tap the wealth of CRISPR possibilities...EsCas13d. Find the Yan et al. plasmids here Find the Konermann et al. Plasmids here How do Cas13d enzymes...VI-D CRISPR Effectors.” Cell (2018) pii: S0092-8674(18)30207-1. PubMed PMID: 29551272 Plasmids from ...Addgene Blog: Learn about CRISPR RNA Editing with REPAIR Get tips for using Cas9 Transcriptional Activators...RNA-editing Cas13 enzymes have taken the CRISPR world by storm. Like RNA interference, these enzymes ...general bioinformatic screens that looked for a CRISPR repeat array near a putative effector nuclease....

  13. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...Find all CRISPR plasmids and resources at Addgene Find CRISPR base editors Find CRISPR plasmids for plants...invented CRISPR-base editors by tethering cytosine deaminase with Cas9 nickase (nCas9). nCas9 creates ...base. If you heard about CRISPR, one question probably comes to mind: can CRISPR be applied to fix the defective...Bastian Minkenberg, and Yinong Yang. "Boosting CRISPR/Cas9 multiplex editing capability with the endogenous...Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion." Nature biotechnology..., Y. Multiplex CRISPR-mediated base editing in rice. In Genome Engineering: The CRISPR-Cas Revolution,...Addgene blog Read all of our CRISPR 101 blog posts Download the CRISPR 101 eBook Read our blog posts ...

  14. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ... Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral Barcoding...and selection marker removal. CRISPR technology pdCas9-humanized (Plasmid #44246) Expression of a catalytically...from the Addgene viral service. CRISPR technology lentiCRISPR v2 (Plasmid #52961) This is a 3rd generation...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling...gamma-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G, may be...expressing new shRNA sequences pLKO (Plasmid #10878) This plasmid has been used by The RNAi Consortium...up-to-date information. Check our CRISPR guide and blog to learn more about CRISPR and the most recent developments...

  15. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast and then selected with...single cells for 13 days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described... Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature...

  16. CasPEDIA: A Functional Classification of Cas Enzymes

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    Blog Post
    ...Addgene blog CRISPR 101: Which Cas9 do I choose for my CRISPR experiment? CRISPR 101: Cas9 The Other Cas...the exact Cas9 plasmid used by Jinek et al. 2012, or browsing for the latest AsCas12a plasmids, Addgene...on Addgene.org CRISPR software and resources Addgene CRISPR Guide Addgene’s CRISPR 101 eBook More ...with any other required non-CRISPR-associated host factors required for CRISPR processing. For each enzyme...Expanding CRISPR Beyond SpCas9 More resources from the Innovative Genomics Institute      CRISPRpedia IGI... gene targeting. Fast forward a decade, and the CRISPR field has exploded with dozens of Cas enzymes and...find a link to search Addgene's collection for plasmids containing that enzyme, and embedded in the References...

  17. CRISPR-Cas14: a family of small DNA-targeting enzymes enabling high-fidelity SNP genotyping

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    Blog Post
    ... to a CRISPR array and the universal CRISPR integrase, cas1. They discovered a family of CRISPR-Cas systems...Addgene.org Find the Cas14 plasmids here Checkout our CRISPR guide page Browse the CRISPR collection ...adjacent CRISPR array and intergenic regions containing the putative tracrRNA into a plasmid and expressed...about DETECTR Browse all of the CRISPR blog posts Download our CRISPR cheat sheet Resources on Addgene.org...genome of virtually any organisms on this planet, CRISPR-Cas systems were merely adaptive immune systems...other Cas proteins found in the so-called class 2 CRISPR-Cas systems. There are 24 variants of the cas14...predicted RuvC nuclease domain, characteristic of CRISPR-Cas enzymes. In contrast to other Cas enzymes, ...

  18. Donations from Addgene to Yield Answers for Rare Disease Researchers

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    Blog Post
    ...induced pluripotent stem cell (iPS) technology and CRISPR-Cas9 constructs from Addgene to gene-edit mutations...looking at a bunch of different strategies with CRISPR-Cas9. There are so many different variants and strategies...role of those mutations, they now plan to use CRISPR-Cas9 technology to create cell lines and/or mouse ...specifically, they’ve proposed to use plasmids with modified nickase Cas9 protein in order to target the specific...studying four of those rare diseases will receive plasmids from Addgene worth a total of $5000.  Lowe Syndrome...involving the primary cilia. They’ll put a variety of plasmids to work to explore the effect of various OCRL1...there. This will help us acquire a lot of those plasmids so that we can try many different things in parallel...

  19. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    ...Editing CRISPR 101: Cas9 Structural Biology Additional resources on Addgene.org Browse CRISPR plasmids...how CRISPR-derived base editing works in general. Base editors use either catalytically dead Cas9 (dCas9...need from Cas9 are sequence specificity and the R-loop structure. In order to bind DNA, Cas9 requires ...to the stretch of ssDNA created by Cas9 binding. Pro tip! In CRISPR, the non-target strand is the DNA ...Acronym Full name Notes CRISPR terms gRNA/sgRNA Guide RNA RNA used by Cas9 to locate a matching DNA...non-target strand of DNA for Cas9 to bind.   R-loop The structure formed by Cas9 binding to DNA and separating... Programmable C:G to G:C genome editing with CRISPR-Cas9-directed base excision repair proteins. Nature...

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

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    Blog Post
    ...endonuclease, Cas9 nickase, Cas9 activators, Cas9 repressors, select gRNAs, and CRISPR pooled libraries....produce CRISPR tools, we’re starting the Viral Service with lentiviral preps for the delivery of Cas9 endonuclease...Service, we’re distributing lentivirus (with many CRISPR tools included among the preps that are currently...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... in experiments other than just those involving CRISPR and so we also have lentivirus for delivering control...
Showing: 101 - 120 of 274 results