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We narrowed to 242 results for: gRNA

Showing: 201 - 220 of 242 results

  1. CRISPR Guide

    Type
    Guide
    ... of unique gRNAs. gRNA design for CRISPR libraries is usually optimized to select for gRNAs with high ... guide RNA ( gRNA or sgRNA ) and a CRISPR-associated endonuclease (Cas enzyme). The gRNA is a short synthetic...commonly referred to as an sgRNA. gRNA scaffold sequence The sequence within the gRNA that is responsible for...amplify a specific sequence of DNA pegRNA P rime E dit gRNA; a specialized gRNA containing an additional primer...Cas enzyme and a gRNA specific to the gene of interest. The genomic target of the gRNA can be any ~20 nucleotide...interactions between the gRNA scaffold and surface-exposed positively-charged grooves on Cas9. gRNA binding induces...if the gRNA spacer sequence shares sufficient homology with the target DNA. Once the Cas9-gRNA complex...

  2. Typing CRISPR Systems

    Type
    Blog Post
    ...Cas12 is unique because it can process multiple gRNAs under a single promoter, allowing for easy multiplexing...

  4. Guide to Using Pooled Libraries

    Type
    Guide
    ...each plasmid contains a unique cDNA. In shRNA or gRNA libraries, each plasmid contains a unique gene targeting...library from the Moffat lab has over 175,000 different gRNA-containing plasmids). Amplifying and Using the Library... screen. The figure above is an outline of how a gRNA library is used. Below are the general steps to ...experimental and control cells Generate a list of gRNAs that disappear with addition of the selection mechanism...

  5. Rett Syndrome

    Type
    Collection
    ... selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of additional CRISPR resources...

  6. Cre-lox system

    Type
    Collection
    ... CMV Lentiviral Lasek 87682 AAV-U6gRNA1-U6gRNA2-TnT-Cre Cre and gRNAs cTnT AAV Pu 87694 pCAG-ERT2-PhoCl-Cre-PhoCl-ERT2... vector Cbh Lentiviral Eisenhoffer 132551 AAV-U6-gRNA-TnT-Cre Cre Chicken cardiac troponin T AAV Pu 134308...60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Cre, luciferase, and sgRNA expression EFS... Zhang 60229 AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR Cre and sgRNA coexpression Cbh AAV Zhang...ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR Cre, KASH-tagged EGFP, and sgRNA expression...expression hSyn AAV Zhang 60820 pSECC Cre, Cas9 and sgRNA expression EF-1 alpha Lentiviral Jacks 60877 pMAZe...

  7. Lentiviral Vector Guide

    Type
    Guide
    ... require gRNAs for each target. Lentiviral vectors are a common way of delivering these gRNAs, due to ...Vector Production Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid ...plasmids: Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs; 8–10 kb packaging...

  8. Addgene's eBook Collection

    Type
    Guide
    ...planning your CRISPR experiment and designing your gRNA and how to use pooled libraries in genome-wide screens...

  9. Adenovirus Guide

    Type
    Guide
    ...Vector Production Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid ...capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single viral particle...

  10. Promoters

    Type
    Guide
    ...also transcribes small RNAs, such as shRNAs and gRNAs. Resources Addgene's blog , including our popular...

  11. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...Vector Production Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid ... Cas9 from Streptococcus pyogenes (SpCas9) and a gRNA together into an rAAV capsid is challenging due ...

  12. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...Vector Production Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid ...Figure 2): Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs; ~8 kb packaging...

  13. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...targeted to their endogenous loci using validated gRNAs from Addgene and well designed homology directed...fused to an sgRNA used to direct Cas9 to a specific genomic locus. When fused to the sgRNA and co-expressed...dCas9 – the dCas9, sgRNA/RNA cargo complex localizes to the sequence targeted by the sgRNA. This allows scientists...machinery that brings the sgRNA/cargo RNA to the appropriate DNA locus. The sgRNA/RNA fusions retained their...construct. The sgRNA targeting Gaussia luciferase can be easily replaced with an sgRNA targeting your ...specific recruitment of multiple copies of VP64 to the sgRNA-targeted gene, resulting in increased activation...SV40, SFFV) or inducible (CMV-TO) cDNA expression, sgRNA (U6) expression, shRNA/shRNA-miR30 constitutive ...

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

    Type
    Blog Post
    ...CRISPRko sgRNA library The Brunello library consists of 77,441 sgRNAs with an average of 4 sgRNAs per gene...contains two sets of sgRNAs; A, the top 3 ranked sgRNAs, and B, the next 3 ranked sgRNAs. Dolcetto was able...containing only 3 sgRNAs (setA) also outperformed CRISPRi libraries with 10 sgRNAs in the sgRNA metrics, confirming...2014 to predict sgRNA activity. In this study scientists examined approximately 1,900 sgRNAs and identified...subsampling analysis of the sgRNAs, Brunello even with only a single sgRNA outperformed another CRISPR...CRISPR library with 6 sgRNAS, highlighting the benefit of sgRNA design in CRISPR libraries. Thus, the Brunello... CRISPRko sgRNA library as pooled library or lentiviral prep! Dolcetto: human CRISPRi sgRNA library While...

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

    Type
    Blog Post
    ...engineered pegRNA” or epegRNA. Although the addition of the RNA pseudoknot sometimes altered epegRNA expression...does just this by including an additional sgRNA. Using this sgRNA, the prime editor nicks the unedited strand...4.0 license. epegRNAs: Structured RNAs with improved stability Traditional sgRNAs are protected from...degraded, the truncated pegRNA can still guide Cas9 and compete with full pegRNAs for access to the target... compared to PEmax with normal pegRNAs, but combining PE7 with epegRNAs resulted in similar or reduced...     ~   3′ RNA pseudoknot (epegRNA) Improve pegRNA stability ~ ~ ~ ~ ~ ~ ✖ ...? First, an engineered prime editing guide RNA (pegRNA) that both specifies the target site and contains...

  16. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...engineered pegRNAs (epegRNAs), which add a structural motif to stabilize the 3’ end of the pegRNA. As research...The team designed sgRNAs that include an internal barcode and separated multiple sgRNAs with tRNAs, which...BFP2.     Figure 1: CROPseq-multi uses two sgRNAs with internal barcodes (iBARs), multiplexed using...lentiviral integration, producing a second copy of the sgRNAs. Image reused from Walton et al. 2024 under a CC-BY-NC-ND...occurs on the RNA polymerase III transcript, the sgRNAs were encoded on the lentiviral minus strand in ...of host cell proteins, and modifications to the pegRNA. The Britt Adamson Lab recently used a CRISPRi ...to La binding and stabilizing the 3’ end of the pegRNA, which is not well protected by Cas9. This is similar...

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

    Type
    Blog Post
    ... activator (CRISPRa) sgRNA library uses the sunCas9 system and contains 10 sgRNAs for each transcription...nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA incorporating two MS2 RNA aptamers at the tetraloop...gain-of-function screening when combined with a compact pooled sgRNA library. Our SAM-mediated screens exhibited a high...transcriptional repressor (CRISPRi) library contains 10 sgRNAs for each transcription start site in those 15,977...further optimize those now. They are also developing sgRNAs that will enable even more fine-tuning of gene ...

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

    Type
    Blog Post
    ...Once supplied with a guide RNA — whether it’s an sgRNA or a crRNA:tracrRNA duplex — Cas9’s two lobes move...yellow; REC lobe, light blue; bridge helix, purple; sgRNA, dark blue.   The REC lobe binds tightly to ...yellow; REC lobe, light blue; bridge helix, purple; sgRNA, dark blue.   DNA sequence recognition Once ...yellow; REC lobe, light blue; bridge helix, purple; sgRNA, dark blue; target DNA strand, light gray; non-target... and marks the transition back to a DNA duplex. sgRNA, dark blue; target DNA strand, light gray; non-target...yellow; REC lobe, light blue; bridge helix, purple; sgRNA, dark blue; target DNA strand, light gray; non-target...

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

    Type
    Blog Post
    ...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...restore the wild-type sequence (save for the residual sgRNAs and possibly cas9 gene). An immunizing reversal...

  20. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...recombination methods). By using mCherry for selection of sgRNA plasmid transformants, they avoid M. abscessus’ ...Using an inducible Cas9 expression and removing the sgRNA plasmid after identifying successful knockouts limits... KanR. This strain is then transformed with the sgRNA cassette on a plasmid expressing mCherry and resistance... of the target gene. Lastly, curing the mCherry-sgRNA plasmid leaves an edited strain carrying Cas9 to...to allow the introduction of additional sgRNAs. Image reused from Neo et al., 2024 under CC-BY license...
Showing: 201 - 220 of 242 results