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  1. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

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    Blog Post
    ...Download Deposit Spreadsheet.” If you’re depositing 10 or more plasmids, the Deposit Spreadsheet is the quickest...and sending the deposit spreadsheet Figure 2: Example of the deposit spreadsheet showing the first eight...Download a new spreadsheet each time you want to submit new plasmids! We update the spreadsheet every so often... more than 10 plasmids is to use our deposit spreadsheet?  Our plasmid deposit process can be broken down...this post, we’ll explain how to use our deposit spreadsheet to complete Step 1. Steps 2 and 3 are usually...send us your plasmid information in a deposit spreadsheet. The data entry process On the Addgene home...Submit Online Deposit” and “Download Deposit Spreadsheet” (see Figure 1). Please read our “How to Deposit...
  2. xCas9: Engineering a CRISPR Variant with PAM Flexibility

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    Blog Post
    ...Check out our CRISPR Guide Find CRISPR Plasmids for Mammalian Systems Learn about CRISPR History ...In order to bind DNA, Cas9 and other CRISPR enzymes require a short PAM sequence adjacent to the targeted...Learn about Engineered Cas9 PAM Variants Read about CRISPR Base Editing Learn about Homology Directed Repair...
  3. CRISPR Challenges: Standardization and Homology Directed Repair

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    Blog Post
    ...Alt-NHEJ), has been harnessed for CRISPR knock-in applications. CRISPR 101: Targeting RNA with Cas13a (...mentioned, John Doench’s How to Design your gRNA for CRISPR Genome Editing, gets over 4,000 views a month. ...design is still challenging for researchers using CRISPR, and that standardizing and improving the gRNA ...necessary going forward. The standardization of CRISPR methods was a common thread at the meeting. The...we need more standardized methods for evaluating CRISPR technologies, including the ever important on/off-target...complementary goals are aimed at improving translation of CRISPR technology, but they would also bring benefits ...terms and techniques are not enough to move the CRISPR field forward. To do that, we need to learn much...
  4. CRISPR 101: Making a Knock-In Cell Line

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    Blog Post
    ...for successful HDR events. CRISPR cut site location Position your CRISPR cut site as close as possible...Efficiency for CRISPR Editing in Human Cells Resources on Addgene.org Browse All CRISPR Plasmids Find...s41589-019-0432-1 Mehdi Banan. Recent advances in CRISPR/Cas9-mediated knock-ins in mammalian cells. J Biotechnol... homology-directed repair choice in response to CRISPR/Cas9 Induced-double strand breaks. Int J Mol Sci...human genome engineering by controlled timing of CRISPR/Cas9 delivery. eLIFE. (2014) 10.7554/eLife.04766...Fernandez, J. P., Rouet, R., Vejnar, C. E., et al. CRISPR-Cpf1 mediates efficient homology-directed repair...Additional Resources on the Addgene Blog Check out our CRISPR Featured Topic Page Learn about Non-Homologous ...
  5. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ...toxicity. More AAV-based CRISPR systems Smaller Cas proteins SaCas9 isn't the only CRISPR enzyme that’s small...about Cas9 Variants and the Expanded CRISPR Toolbox Read about CRISPR enzyme Cpf1 Read our Introduction ...Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids ... and last updated Sep 16, 2020 by Beth Kenkel.  CRISPR genome editing has quickly become a popular system...characterization for in vivo studies. One of the pitfalls of CRISPR/Cas9 genome editing is the potential for off-target...liver-specific serotype AAV8. Since the efficiency of CRISPR/Cas9 genome editing varies across targets, they...added advantage that it can be turned off by anti-CRISPR proteins. The Sontheimer lab used NmeCas9 with ...
  6. The PAM Requirement and Expanding CRISPR Beyond SpCas9

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    Blog Post
    ...same species. Expanding the CRISPR toolbox The isolation of novel CRISPR proteins has and will continue...dramatically increase the number of CRISPR applications. The first non-Cas9 CRISPR protein adapted for genome ...trying to decide which CRISPR tools are best for your experiment, check out our CRISPR guide or leave a comment...Check out our CRISPR Featured Topic Page Resources on Addgene.org Check out our CRISPR Guide Browse ...variants effectively double the targeting range of CRISPR/Cas9 within the human genome. Another variant, ...near-PAMless Cas9 variants. These variants, named SpG and SpRY, can target NGN PAMs and NRN and NYN PAMs, respectively...AT-rich genomes (Kleinstiver et al., 2016). Type VI CRISPR systems, which target RNA, offer additional targeting...
  7. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...blog Browse our CRISPR blog posts Download our CRISPR cheat sheet Download our CRISPR eBook Resources...out our comprehensive CRISPR guide Find CRISPR plasmids and resources Find CRISPRa plasmids ...recent papers describe the development of CRISPR activation (CRISPRa) tools for neuroscience, including transgenic...neuroscience research. What is CRISPR-based activation? To summarize, CRISPRa uses a nuclease-dead variant...by specific sgRNAs. Its RNA-guided nature makes CRISPRa much simpler to implement in genome-wide screens...screens over cDNA libraries. However, using CRISPRa to target individual genes can be relatively laborious...ones. Simplified in vivo delivery using SPH mice CRISPRa has largely been limited to in vitro settings due...
  8. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...blog Download the CRISPR 101 eBook Find our CRISPR 101 blog posts Read our CRISPR cheat sheet Resources...Resources on Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids...While much of CRISPR research has focused on genome editing, numerous discoveries have been made using...genomic region of interest targeted by a gRNA. CRISPR-mediated purification of a specific genomic region...method to purify a specific genomic region using the CRISPR system, consisting of dCas9 and targeting gRNA,...require multiple cloning steps (Fujita et al., 2013). CRISPR-mediated purification of a specific genomic region...Experimental use Since the initial report describing CRISPR-mediated purification of a specific genomic region...
  9. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...Find more CRISPR Resources at Addgene: Read our CRISPRGuide Get practical help from our CRISPR Protocols...Tools Read other CRISPR blog posts Find CRISPR Plasmids at Addgene  ...realistic. Check Out Our Post on Sequencing Options for CRISPR Genotyping NGS and detecting off target effects... to an untreated population. Software such as CRISPResso can help with the data analysis. If NGS is not.... Luckily, in silico prediction analyses like CRISPRitz or CRISPOR can help narrow down the list of sites...Steensel. Easy quantification of template-directed CRISPR-Cas9 editing. (2018) Nucleic Acids Research Volume..., Canver MC, Bombieri N, Giugno R, Pinello L. CRISPRitz: rapid, high-throughput and variant-aware in silico...
Showing: 71 - 80 of 559 results