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We narrowed to 94 results for: sgrna

Showing: 21 - 40 of 94 results

  1. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...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 ...

  2. 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...

  3. CRISPR References and Information

    Type
    Collection
    ... Sabatini and Lander gRNA cloning into pLX-sgRNA pLX-sgRNA PDF, 129 KB Sontheimer gRNA design and cloning... can identify positively or negatively selected sgRNAs, genes, or pathways. For use with genome-scale ... opens in a new window) for lentiviral screens. sgRNA Scorer 2.0 (Link opens in a new window) A tool that... and a ctivation (Link opens in a new window) A sgRNA design tool for genome editing, as well as gene ...Link opens in a new window) Identifies candidate sgRNA target sites by off-target quality. Validated for... (Link opens in a new window) Identification of sgRNA target sites and off-target evaluation for base ... window) Ranks and picks candidate CRISPRko/a/i sgRNA sequences to maximize on-target activity for selected...

  4. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...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 ...and shRNAs with a reporter gene, or (4) multiple sgRNAs with Cas9 for genome engineering. The versatility...

  5. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...indicating loci where SNPs reside on sgRNA target sequences for sgRNAs that target ancestry-associated genes...that could analyze CRISPR sgRNAs and report the likelihood of any given sgRNA having an ancestral bias....

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

    Type
    Blog Post
    ...activity coupled with a customizable single guide RNA (sgRNA) that bears complementarity to a 20bp target sequence...multidirectional control, a new study repurposes the sgRNA to include modular synthetic RNA scaffolds that ...modules are comparatively small allowing multiple sgRNA constructs to be introduced simultaneously in a ...

  7. CRISPR Pooled gRNA Libraries

    Type
    Collection
    ...46,354 BARBEKO sgRNA Library 174163 Knockout, Base Editing Human Wei 3rd 3 53,502 sgRNA Pool for Streptococcus...131625 Inhibition E. coli Bikard N/A ∼5 21,417 Bison sgRNA Library 169942 Knockout Human Ebert 3rd 4 2,852 ...Fly Perrimon 3rd 8 6 6 7,956 17,827 59,406 Dual sgRNA CRISPRi Libraries 187246 — 187248 1000000192 Inhibition...Inhibition Human Weissman 3rd 6 Varies Dual sgRNA CRISPRa Libraries 187249 — 187251 1000000193 Activation...3rd 2 37,722 Green monkey (Chlorocebus sabaeus) sgRNA library 178284 Knockout Green Monkey Doench and ...Knockout Human Parts 3rd 3 59,576 Human Whole Genome sgRNA iBAR Library 140633 Knockout Human Wei 3rd 3 58,... Mouse Knouse 3rd ~5 71,878 Mouse messenger-RBP sgRNA Library 169082 Knockout Mouse Turner N/A 10 13,500...

  8. 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...

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

    Type
    Blog Post
    ...complexed with sgRNA. Direct comparison of Cas9 RNPs prepared with an in vitro-transcribed (IVT) sgRNA versus ...cumbersome in vitro synthesis of single guide RNAs (sgRNAs), are time-consuming, have been typically optimised... the in vitro-transcribed single guide RNA (IVT sgRNA) approach (Dewari et al. 2018 and Figure 2). Not...construct plasmid vectors or synthesize/purify IVT sgRNA in the lab. Figure 2: Cas9 RNP complexed with 2...

  10. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...assemble multiple unique sgRNAs into a multi-TU targeting construct (Cas9, sgRNA, reporter, etc.). This ...for the multiplexing of sgRNAs against unique genomic targets or multiple sgRNAs against the same target...target to ensure efficient cutting. Beyond sgRNAs, MoClo can also be used to test a variety of Cas enzymes...

  12. Hot Plasmids February 2024

    Type
    Blog Post
    ...endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in human cell...limited (likely due to its long, negatively charged sgRNA), RNP delivery was much better with Cas12, which...Workflow to assess Cas9-PAGE editing with viral sgRNA targeting mCherry reporter (mChe), with co-incubation...

  13. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...around 20 base pairs long. Using a single guide RNA (sgRNA) to create a double-strand break works well because...locus (Ho et al. 2012). For large-scale screening, sgRNA libraries are useful for targeting thousands of ...uses a nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily block the transcription machinery...

  14. CRISPR Plasmids - C. elegans

    Type
    Collection
    ... (PAM = NGG) 47549 pDD162 (Peft-3::Cas9 + Empty sgRNA) R07E5.16 U6 yes, cut Goldstein 42250 DR274 T7 BsaI...transcription none, need Cas9 plasmid Boxem 47912 SP6-sgRNA-scaffold SP6 AflII In vitro transcription none, ...need Cas9 plasmid Sternberg 177783 L4440_BioBrick-sgRNA T7 BbsI, BsaI Ingestion by C. elegans none, need...transcription none, need Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9...

  16. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...Zeocin®/Bleo Mammalian lenti sgRNA(MS2)_zeo backbone - 3rd gen lenti sgRNA cloning backbone with MS2 loops...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 expression plasmid for two sgRNAs from an hU6 and an mU6 promoter Protein-protein interaction...confers hygromycin resistance pRGEB32 - Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice...3rd gen lentiviral vector lentiSAMv2 - Lentiviral sgRNA cloning backbone pLKO.1-blast - 3rd gen lentiviral...

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

    Type
    Blog Post
    ...does just this by including an additional sgRNA. Using this sgRNA, the prime editor nicks the unedited strand...increases indel formation slightly. Designing the sgRNA with a spacer that only binds the edited strand,...Structured RNAs with improved stability Traditional sgRNAs are protected from cellular degradation by Cas9...efficiency ✓ ✓ ✓ ✓ ✓   ✓ Second sgRNA to guide nicking of the unedited strand Favor ...

  19. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    ...research with a simple question: how do Cas9 and sgRNA interact with and dissociate from a DNA target? ..., requiring expressed Cas9 protein, transcribed sgRNA, and optional donor DNA. Chris Richardson is a...

  20. Components of CRISPR/Cas9

    Type
    Blog Post
    ...  The synthetic guide RNA or gRNA (sometimes sgRNA) In the native Type II CRISPR/Cas system, Cas9 is...known as the guide RNA (gRNA) or single guide RNA (sgRNA). Throughout the remainder of this post, we’ll refer...
Showing: 21 - 40 of 94 results