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We narrowed to 108 results for: cas9 variant

Showing: 81 - 100 of 108 results

  1. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...Addgene. CRISPR/Cas9 Delivery rAAV vectors are a popular choice for the delivery of CRISPR/Cas9 elements, making... genome editing. Packaging the Cas9 from Streptococcus pyogenes (SpCas9) and a gRNA together into an rAAV...this obstacle, has been to use smaller Cas9 orthologs, such as SaCas9 from Staphylococcus aureus . The Feng...vectors. AAV Types Serotypes AAV serotypes are AAV variants that differ in their tropism — that is, the types...vectors for in vivo applications. Other Engineered Variants Hybrid capsids Hybrid capsids are engineered capsids...successfully employed to engineer new rAAV capsid variants, such as systemic capsids. A systemic capsid is...also been employed to engineer an rAAV serotype variant able to display neuronal retrograde transport, ...

  2. Human Germline Editing Using CRISPR

    Type
    Blog Post
    ... samples. Ma et al. designed plasmids containing Cas9 and two different gRNAs, and electroporated them...genome sequencing in selected control and CRISPR-Cas9 injected embryos. In all cases, they did not observe...CRISPR Topic Page Read about High Fidelity CRISPR Variants Target Multiple Genes at Once with CRISPR Resources...

  3. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...using CRISPR/Cas9 to edit a gene, the expression of Cas9 is no longer necessary. However, Cas9 can be toxic...like creating stable cell lines, delivering CRISPR/Cas9 to cells for genome editing, and introducing inducible...Leavitt et al., 1996), and since then, numerous variants have been developed with different mechanisms ...

  4. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...2010 Yuhai Cui Red mScarlet Split-wrmScarlet variant of mScarlet Split-wrmScarlet and split-sfGFP: tools...-10) and sfCherry2(11), also photo-activatable variant PAsfCherry2(1-10) Improved split fluorescent proteins...Split-horseradish peroxidase (Alice Ting Lab 2016)  Split-Cas9 for inducible genome editing (Feng Zhang Lab 2015...

  5. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...Blainey’s lab has developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi...Anzalone et al., 2019, including modifications to the Cas9 or reverse transcriptase enzymes, inhibition of ...end of the pegRNA, which is not well protected by Cas9. This is similar to the proposed mechanism for engineered...domain conferred several new improvements. The new variant has increased signal-to-noise ratio and better ...of applications, including CRISPR activation by dCas9-VPR, prime editing, and delivery of disease-relevant...

  6. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ... line knock-ins of patient mutations which are variants of unknown significance – mutations with unclear...CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease Out of the genome Tips for a first...

  7. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ... mutants, identifying the two  best-performing variants as iGluSnFR4f (fast deactivation) and iGluSnFR4s...Bentley For RNA-guided genome editing, think beyond Cas9 with STITCHR: a combined CRISPR / retrotransposon.... Figure 4: STITCHR consists of a nicking Cas9 fused to a shortened R2Tocc transposon protein, ...for scarless insertion of the payload (teal). The Cas9 gRNA targeting guide and the transposon RNA homology...scientists fused the retrotransposon protein R2Tocc to a Cas9 nickase to improve target recruitment and nicking...TurboCas provides the genomic targeting power of dCas9 with the dynamic labeling power of miniTurboID, ...proximity ligation tool (Cenik et al., 2024). While dCas9 has been used in combination with proximity labeling...

  8. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...blog CRISPR 101: Which Cas9 do I choose for my CRISPR experiment? CRISPR 101: Cas9 The Other Cas(s) CRISPR...more specific variants. HypaCas9, sSpCas9(1.1), SpCas9HF1, and evoCas9 are a few spCas9s which have been...Ma X, Gao F, Guo Y. Off-target effects in CRISPR/Cas9 gene editing. Front Bioeng Biotechnol. 2023 Mar ...of Maya Kostman. The most commonly used Cas, spCas9, has gone through multiple rounds of optimization...

  9. Validated gRNA Sequences

    Type
    Collection
    ...a single species or PAM binding variant of Cas9 (e.g. Wild-type SpCas9 must be used with targets that ...activity. Know your target. Which species or variant of Cas9 ( S. pyogenes, S. aureus etc.) was this gRNA...C. elegans TCCGCTACCATAGGCACCA 71479 cut VQR Cas9 variant 26680661 Fire dpy‐10 C. elegans GCTACCATAGGCACCACGAG...and Resources page have been used to indicate the Cas9 application the gRNA was designed to accomplish....Target Species Target Sequence Plasmid ID Application Cas9 Species PubMed ID Depositor OCT4 H. sapiens CTCCCATGCATTCAAACTG...10 C. elegans CGCTACCATAGGCACCACG 71516 cut VRER Cas9 mutant 26680661 Fire dpy-10 C.elegans GCTACCATAGGCACCACGAG...Christiaen EGFP A. victoria multiple, see article 60071 dCas9-FokI S. pyogenes 24770325 Joung EGFP A. victoria...

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

    Type
    Blog Post
    ...) model consists of a catalytically dead Cas9 protein (dCas9) that lacks endonuclease activity coupled...effectively change metabolic flux. The dCas9 protein and orthogonal variants can be used as regulatory ON/OFF.... Using dCas9 as an on/off switch The multiple scRNAs all individually utilize the same dCas9 protein....modulate dCas9 expression, multiple scRNA transcriptional programs can be regulated with dCas9 as a master...an orthogonal dCas9 protein that will still recognize guide RNA but with a different dCas9 binding module...specific proteins via aptamer domains. Combined with dCas9, this new scaffold RNA (scRNA) will bind to a PAM-adjacent...layered with distinct programs controlled by each dCas9 regulatory switch. By no means limited to the field...

  11. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...from single or multiplex CRISPR-Cas9 double-strand DNA breaks using SpCas9. More features will be added ...Currently, ICE is configured for Cas9 fromStreptococcus pyogenes (SpCas9). ICE Score - The editing efficiency...Browse our CRISPR featured topic page xCas9: Engineering a CRISPR Variant with Pam Flexibility RNA Editing ...

  12. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...High fidelity Cas9s: Three high fidelity Cas9 variants , SpCas9-HF, eSpCas9, and HypaCas9 display very ...III and Cas9 enzymes, which together separate the individual crRNAs. (3) The tracrRNA and Cas9 nuclease...Staphylococcus aureus (SaCas9) is smaller than SpCas9 and more easily packaged in AAV. Cas9 orthologs also have...beyond SpCas9, moving us closer to being able to target every locus in every genome. Multiple Cas9 orthologs...capability of Cas9 to make other types of modifications at specific loci. Activate or Repress : dCas9 fused to...the crRNA would bind. (5) After the complex binds, Cas9 separates the double stranded DNA target and cleaves...Single multi-domain effector) Type I (Cas3) Type II (Cas9) Type III (Cas10) Type V (Cpf1) Type IV (Csf1) Type...

  13. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    ...condition that can apply enough pressure to select for variants of interest, though. A selection can take the ... Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166): 80-84 (2014). doi: https...

  14. Plan Your Experiment

    Type
    Guide
    ...). You will decide how to express Cas9, the delivery system for Cas9 and the guide RNA (gRNA), the design...non-homologous end joining (NHEJ) at the cut site Cas9 or Cas9 nickase High-fidelity Cas enzymes increase specificity...insertions, by copying from a DNA donor template Cas9 or Cas9 fusions with proteins that promote HDR Requires...site. Consider alternative Cas9 enzymes with different PAM sequences or Cas9 enzymes with flexible PAM .../or Cas9 expression or (2) a lack of efficient target cleavage in cells that do express both Cas9 and ...edits without double-strand breaks Base editor (dCas9 or Cas9 nickase fusions with different base editing ...We will focus on making single edits using CRISPR/Cas9 in mammalian cells as an example, but many of these...

  15. Plan Your Experiment

    Type
    Collection
    ...). You will decide how to express Cas9, the delivery system for Cas9 and the guide RNA (gRNA), the design...non-homologous end joining (NHEJ) at the cut site Cas9 or Cas9 nickase High-fidelity Cas enzymes increase specificity...insertions, by copying from a DNA donor template Cas9 or Cas9 fusions with proteins that promote HDR Requires...site. Consider alternative Cas9 enzymes with different PAM sequences or Cas9 enzymes with flexible PAM .../or Cas9 expression or (2) a lack of efficient target cleavage in cells that do express both Cas9 and ...edits without double-strand breaks Base editor (dCas9 or Cas9 nickase fusions with different base editing ...We will focus on making single edits using CRISPR/Cas9 in mammalian cells as an example, but many of these...

  16. Hot Plasmids - August 2020

    Type
    Blog Post
    ...utility of the system for use with low affinity variants of TEV. The new system has been demonstrated to...the evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription...fluorescence protein-based calcium sensors (jYCaMP1), variants of the calcium indicator jGCaMP7, that are optimized...new calcium sensors include syn-jYCaMP1s (slow variant), the Cre-dependent syn-FLEX-jYCaMP1s, and the ...an EGFP reporter. Expression of the dCas9-KRAB activator or dCas9-VPR repressor in hPSCs is also driven...piggyBac gRNA delivery and the all-in-one dCas9-KRAB and dCas9-VPR targeting plasmids can be found at Addgene...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly...

  17. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...inserted the deaminase into the C-terminal domain of Cas9, which narrowed the editing window and reduced A...site. Like in most other base editor designs, the Cas9 nicks the non-edited strand to encourage the cell...models-assisted optimization of a uracil-N-glycosylase variant enables programmable T-to-G and T-to-C base editing.... gGBE consists only of engineered MPG fused to nCas9. First, MPG removes the G base from DNA to create...

  18. The 12 Days of CRISPR

    Type
    Blog Post
    ...CRISPR, Addgene gave to me: Functional Genetic Variants Revealed by Massively Parallel Precise Genome ...CRISPR, Addgene gave to me: High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral...🎵 On the 3rd day of CRISPR, Addgene gave to me: dCas9-based gene editing for cleavage-free genomic knock-in...

  19. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...recombinase variants to optimize PASSIGE efficiency (Pandey et al., 2024). The best-performing variant yielded...TALEs), with fifteen total variants deposited with Addgene. These variants, including a “miniCRISPR” version...allowing the team to develop self-silencing CHARM variants that stop their own expression after silencing... a programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase — that can drive...DNA-targeting proteins, so the team created versions using dCas9, zinc finger proteins, and transcription activator-like... version, are much smaller proteins than SpyCas9, allowing them to be packaged in AAV vectors, potentially...histone tail (H3K4me0) to a DNA-binding protein (dCas9 shown here). D3L recruits the endogenous, autoinhibited...

  20. Cre-Lox and Other Site-Specific Recombinases

    Type
    Collection
    ...Site Canonical Sequence Structure (Length) Common Variants Cre loxP ATAACTTCGTATAgcatacatTATACGAAGTTAT 13...results. Attenuated iCre(R297T), a point mutant iCre variant, reduces the number of recombination events but...vivo using a mouse line expressing Cre-dependent Cas9. Kits Kit Name Type PI Description dRMCE Plasmid...
Showing: 81 - 100 of 108 results