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We narrowed to 235 results for: crispr grnas

Showing: 201 - 220 of 235 results

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

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    Blog Post
    ... the CRISPR system was a large one.  The question for her then was this: Can you engineer CRISPR-Cas9 ...Two-way CRISPR screening In October, Jonathan Weissman’s lab introduced additional pooled CRISPR libraries...CRISPR technology has changed how scientists edit and control genes, but according to the Broad Institute's...Institute's Silvana Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation...lab describe just such a system, which they call CRISPR/Cas9 Synergistic Activation Mediator or SAM. SAM...earlier by the Zhang lab for GeCKO, a lentiviral CRISPR library enabling genome-scale, knockout screening...review the steps for how to use pooled lentiviral CRISPR libraries and keep the following in mind: These...

  2. Analyzing CRISPR Editing Results with ICE from Synthego

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    Blog Post
    ... Check out our CRISPR guide Find CRISPR plasmids for your research Find other CRISPR software resources... Guide for Analyzing CRISPR Editing Results with ICE on Synthego’s blog. CRISPR-based genome engineering...Inference of CRISPR Edits) to solve this problem. This tool was initially created to support the CRISPR analysis...successful CRISPR experiments. Jessica is a sci-fi enthusiast, and when she is not guiding CRISPR researchers...Addgene Blog Browse our CRISPR featured topic page xCas9: Engineering a CRISPR Variant with Pam Flexibility...methods. Still, generating reliable results from CRISPR edit data requires the help of robust software ...to produce quantitative, NGS-quality analysis of CRISPR editing - enabling a ~100-fold reduction in cost...

  3. Uncovering Ancestral Bias in CRISPR Research

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    Blog Post
    ...tool that could analyze CRISPR sgRNAs and report the likelihood of any given sgRNA having an ancestral bias...extension, race. Many researchers do large-scale CRISPR screens in cancer cell lines, looking to uncover... and Boehm labs pulled data from more than 1000 CRISPR screens in DepMap to see if they could identify...Since about 90% of the cell lines used in DepMap CRISPR screens are of either European or East Asian ancestry...it was coming through an unexpected source: the CRISPR guides, short pieces of RNA that bind to a cell...does the reference genome actually reference?  The CRISPR guides used in the screens they analyzed were designed...reference genome – even single mismatches within a CRISPR guide can prevent the guide from binding to the...

  4. The 12 Days of CRISPR: 2021

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    Blog Post
    ...Twelve Days of CRISPR campaign! But if you missed our tweets featuring the best of CRISPR in 2021 and useful...here.  On the first day of CRISPR, Addgene gave to me: a miniature CRISPR-Cas system for mammalian use...machinery. On the fifth day of CRISPR, Addgene gave to me: efficient CRISPR editing with a hypercompact ...seventh day of CRISPR, Addgene gave to me: programmed genome editing by a miniature CRISPR-Cas12f nuclease... day of CRISPR, Addgene gave to me: a primer on prime editing.  On the eleventh day of CRISPR, Addgene...post.  On the twelfth day of CRISPR, Addgene gave to me: an updated CRISPR 101 e-book (to help with the... get enough of CRISPR? Neither can we! Subscribe below to get updates on all our CRISPR content.  ...

  5. A Needle in a Base-Stack: Cas9 Structural Biology

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    Blog Post
    ...Addgene's CRISPR Guide Addgene's CRISPR 101 eBook  Resources on the Addgene blog Components of CRISPR  xCas9...engineered CRISPR enzymes with their own tradeoffs. And this article only covered Cas9! Other CRISPR enzymes...Have you ever designed a CRISPR guide RNA and wondered why it is limited to only 20 bases, or why it’...structure can help us understand the parameters of CRISPR experimental design. Let’s dive into some structural...known proteins; they are completely unique to the CRISPR system (Nishimasu et al., 2014). Table 1. Summary...positively charged channel between the two lobes. For CRISPR to be programmable for any arbitrary sequence, ...better understand how to manipulate it. Happy CRISPRing!   Molecular graphics and analyses performed with...

  6. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...10.1186/s12915-020-00929-7    The CRISPR corner By Jennifer Tsang New CRISPR plasmids are always being added...To find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources...other pneumococcal strains. The BARBEKO sgRNA library uses CRISPR cytosine base editors to disrupt gene ...A new tool for enriching ABE-targeted cells The CRISPR Corner New ready-to-use viral preps   Improved..., in addition to puromycin resistance.  The MYC-CRISPR library targets E-boxes genome-wide. It was designed...involved in DNA damage response and DNA repair.  A new CRISPR pooled library targeting S. pneumoniae D39V from...Veening is now available with one sgRNA per operon or gene. The sgRNA pool can also target core operons...

  7. Using CRISPR/Cas9 to Edit Disease Out of the Genome

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    Blog Post
    ...post on CRISPR’s potential for monogenetic diseases, the bulk of which you can find below. CRISPR for complex...proof-of-principle for use of CRISPR to correct single-gene conditions. Remaining hurdles in CRISPR biology “I think...using sgRNA for selection and optimization. In addition, the current work in developing anti-CRISPR proteins...Northeastern University.  There can be no doubt that CRISPR/Cas9 technology has been a breakthrough for the... complex genetic diseases As CRISPR technology becomes increasingly more cost effective and precise, researchers...diseases. So we’re revisiting this post to look at CRISPR’s potential for use in treating a complex genetic...his lab are researching a way to treat AD using CRISPR/Cas9. They are targeting the genes responsible ...

  8. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...can bind loci in the genome that are defined by a gRNA. Since Cas9 is a protein, a fusion with GFP allows...genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7 (2013): 1479-1491. PubMed...

  9. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ...Guide RNAs Visit our CRISPR topic page Learn the basics of CRISPR Download the CRISPR 101 eBook Resources...Resources on Addgene.org Read the CRISPR guide Find CRISPR plasmids by function Browse our entire plasmid ...variants in most cell types. But prime editing, a CRISPR technique developed in David Liu’s lab in 2019,...considerably more precision and flexibility to the CRISPR editing world. The approach, originally published... location The prime editor extends the reach of CRISPR genome editing, as it can edit near to or far from...brief discussion of some of these strategies in our CRISPR guide. Some examples include: TwinPE: two prime...sequences without double-strand DNA cleavage using CRISPR-directed integrases. Nature Biotechnology, 41(4...

  10. Empty Backbones - Choosing Your Perfect Plasmid Backbone

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    Collection
    ...plants Visit our CRISPR Plasmids and Resources collection, and browse our Empty gRNA Expression Vectors... Gateway compatible Y2H vector - BD pRPR1_gRNA_handle_RPR1t - gRNA expression empty vector pFA6a vectors...Representative Empty Backbones CRISPR Genome modification lentiCRISPR v2 - 3rd gen lentiviral vector ...selectable marker pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP - For retroviral delivery of one sgRNA pMKO.1 GFP - Retroviral...- Deliver sgRNA and Cas9 into plants by Agrobacterium-mediated transformation Dual-sgRNA_hU6-mU6 - Lentiviral... Lentiviral shRNA expression lentiCRISPRv2 hygro - Variant of lentiCRISPR v2 that confers hygromycin resistance...Zeocin®/Bleo Mammalian lenti sgRNA(MS2)_zeo backbone - 3rd gen lenti sgRNA cloning backbone with MS2 loops...

  11. Plan Your Experiment

    Type
    Guide
    ...how to design your gRNA . Browse Plasmids: gRNAs Synthesize and clone desired gRNAs Once you have selected... Science Guides CRISPR Guide Plan Your Experiment Plan Your CRISPR Experiment Jump To: Plan Your Experiment... Select Expression System Design gRNA Publications Get Started CRISPR is a powerful system that enables...you started using CRISPR in your research. Although we will use the example of CRISPR/Cas9 in mammalian...Select Expression System To use CRISPR, you will need both Cas9 and a gRNA expressed in your target cells...express the CRISPR machinery. For more difficult cells (e.g. primary cells), viral delivery of CRISPR reagents... using CRISPR in mammalian cells. Some of the variables include: Species of Cas enzyme and gRNA Species...

  12. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...Modifying the CRISPRi system for multidirectional control The CRISPR interference (CRISPRi) model consists...Addgene: Plasmids for CRISPRi are described here: http://www.addgene.org/crispr/qi/ Plasmids described...Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds." Adam has also created a video to...metabolic biofactories by demonstrating the use of CRISPR-based RNA scaffolds to mimic natural transcriptional...AdamChinFatt     References Qi, L. S. et al. Repurposing CRISPR as an RNA-guided platform for sequence-specific...Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell, 160(1), 339-350 (2015). PubMed... crispr...

  13. Hot Plasmids: Fall 2024

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    Blog Post
    ...additional sgRNAs. Image reused from Neo et al., 2024 under CC-BY license.   Find plasmids for CRISPR/Cas9...mScarlet-based tags for live imaging in C. elegans CRISPR/Cas9 for gene disruption in M. abscessus  Bioluminescent...in Mycobacterium abscessus using a dual-plasmid CRISPR/Cas9 system By Aryana Khosravani Deborah Hung’s...’s lab has developed a novel dual-plasmid CRISPR/Cas9 system for gene disruption in Mycobacterium abscessus...for its high resistance to antibiotics. Several CRISPR/Cas-based systems have been developed for other...errors (Figure 4). This work marks the first use of CRISPR/Cas9 in M. abscessus. The simplicity and efficiency...   Figure 4. Overview of the dual-plasmid CRISPR/Cas9 workflow in M. abscessus. A) Sth1Cas9 mediates...

  14. CRISPR Guide

    Type
    Guide
    ... of unique gRNAs. gRNA design for CRISPR libraries is usually optimized to select for gRNAs with high ... Addgene Blog CRISPR topic page How to design your gRNA for CRISPR genome editing CRISPR 101: Homology...basic CRISPR mechanism Engineered CRISPR systems contain two components: a guide RNA ( gRNA or sgRNA ) and...Guide CRISPR Guide Jump To: CRISPR Guide Overview CRISPR Basics Small Edits Large Edits Genome-Wide Screening...generating gRNAs makes CRISPR one of the most scalable genome editing technologies, making CRISPR perfect...Figure 7: Overview of CRISPR transposases Browse Plasmids: CRISPR transposases CRISPR Recombinases and Integrases...screening using CRISPR libraries What Are Pooled Lentiviral CRISPR Libraries? Pooled lentiviral CRISPR libraries...

  15. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...pooled CRISPR screens By Emily Bentley Building on their CROPseq method for optical pooled CRISPR screening...Multiplexed perturbation and decoding in pooled CRISPR screens Improved prime editor PE7 Antibodies to...developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi (Walton et al. 2024...multiplexed perturbation and decoding in pooled CRISPR screens. bioRxiv 2024.03.17.585235; doi: https:...sequences for a range of applications, including CRISPR activation by dCas9-VPR, prime editing, and delivery...including plasmids, antibodies, viral vector preps, CRISPR tools, and more.  ...engineered pegRNAs (epegRNAs), which add a structural motif to stabilize the 3’ end of the pegRNA. As research...

  16. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    ...summer of 2013, a remarkable nine papers describing CRISPR/Cas9 genome engineering methods for C. elegans ...lab at the University of North Carolina has made CRISPR in C. elegans even easier - now, one can generate...approach Multiple features of C. elegans have made CRISPR/Cas9 so successful in this species, namely, the...transgene selection strategy for their C. elegans/CRISPR experiments, but this strategy required the use...sought to create a new and efficient selection-based CRISPR/Cas9 method to generate knock-ins on any genetic...other species or in cell culture. In addition to CRISPR/Cas9 applications, SEC selection could replace ... system represents another leap forward for the CRISPR/Cas9 revolution!   References Streamlined Genome...

  17. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...Nature Methods. 2015 Aug 31. CRISPR-Display: A novel CRISPR technology for targeting RNA cargos...technique, called CRISPR-Display, that may help answer this difficult question. CRISPR-Display uses the...specific DNA loci. In CRISPR-Display, your RNA of interest is fused to an sgRNA used to direct Cas9 to...system provides users greater temporal control over CRISPR/Cas9 mediated genome modification and gene expression...139-42. doi: 10.1038/nbt.3149. Light-inducible CRISPR-Cas9 system for endogenous gene activation Optogenetics...Nucleases, the Gersbach lab has modified the RNA-guided CRISPR-Cas9 system to create a tool that is quick, versatile...robust. Dubbed the LACE system for light-activated CRISPR-Cas9 effector, Polstein and Gersbach fused the ...

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

    Type
    Blog Post
    ....org Read our CRISPR guide Find CRISPR plasmids for your research Find links to CRISPR references ... also led to the discovery of IgnaviCas9, a new CRISPR enzyme from a sample from Mound Spring in Lower...throughput and rigorous assembly. There, they found a CRISPR array in the resulting Ignavibacteriae phylum genome... genome. IgnaviCas9 is a hyperthermostable Cas9 CRISPR nuclease that is active up to 100°C.  The IgnaviCas9...temperatures for long time periods and workflows where CRISPR nuclease activity is desired and time or protocol... Learn about other Cas proteins Read all of our CRISPR blog posts Read about GeoCas9, another thermostable... protein is 1,241 amino acids long, has optimal sgRNA spacer length of 22-25 nts, and is thermostable ...

  19. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ... the ZFNs.   Genetic Engineering Era: Post CRISPR  CRISPR The strategy of using a protein to recognize... Find more blog posts about CRISPR Learn more about the history of CRISPR Read about other genome editing...methods Resources on Addgene.org Visit our CRISPR guide Find CRISPR plasmids for your research Browse plasmids...during this period.  Genetic Engineering Era: Pre-CRISPR  Having visualized and understood the DNA, the ...breakthrough in the world of genetic engineering as CRISPR-Cas editing soon replaced the protein-based DNA-binding... gene editors (meganucleases, ZFNs, TALENs, and CRISPR-Cas) is that they induce double-strand breaks (...variations in the genome.    Due to the rise of CRISPR in genetic engineering and open advertisement of...

  20. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...PubMed. More CRISPR-Cas9 Resources: CRISPR 101: a blog series on the basics of CRISPR-Cas technology...technology CRISPR Resources from Addgene   ... crispr...cassette encoding the cas9 gene and single guide RNAs (sgRNAs) with appropriate flanking homology into a germline...don't use a DNA vector that encodes both Cas9 and sgRNA. What if I want to work with gene drives? Then do...organisms but not wild organisms. For example, an sgRNA-only drive will spread exclusively through populations...express Cas9 from an unlinked locus, while a Cas9+sgRNA drive targeting a synthetic sequence will only spread...
Showing: 201 - 220 of 235 results