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We narrowed to 110 results for: cas9 variant

Showing: 81 - 100 of 110 results

  1. 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...

  2. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...editors use either catalytically dead Cas9 (dCas9) or a Cas9 nickase (nCas9) to avoid introducing double-strand...need from Cas9 are sequence specificity and the R-loop structure. In order to bind DNA, Cas9 requires ...expanded CGBE target scope by creating BE variants with other Cas9s that recognize different PAMs. Their computational...non-target strand of DNA for Cas9 to bind.   R-loop The structure formed by Cas9 binding to DNA and separating...PAM) on the opposite strand. Once it finds a PAM, Cas9 unwinds the adjacent DNA to create a region of separate...single-stranded DNA (ssDNA) region, using enzymes fused to Cas9 to alter the exposed ssDNA bases. Thus, base editors...window restricted to the stretch of ssDNA created by Cas9 binding. Pro tip! In CRISPR, the non-target strand...

  3. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ... third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors. Effector...-based biosensor for cAMP dynamics. The mGold2 variants significantly enhanced photostability — 25-fold..., recruited by either the ωRNA (TnpB) or a gRNA (Cas9). Drawings not to scale. Created with BioRender....

  4. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ... plasmid contains Cas9 and if so, which function of Cas9. We have plasmids with Cas9 that can cut, activate...Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI pCRISPR 42875 Bacteria BsaI...BbsI)_CBh-Cas9-T2A-BFP 64323 Mammalian BbsI yes, cut S. pyogenes BFP Kuhn pU6-(BbsI)_CBh-Cas9-T2A-mCherry...meningitidis Joung Cas9 sgRNA vector 68463 Mammalian U6 none S. pyogenes Zhang Cas9/pTREX-n 68708 Other...independent U6 promoters and Cas9 from CBh promoter. Ventura A CRISPR/Cas9 toolkit for multiplex genome...cassettes are assembled into a Cas9 expressing vector. Yang A Robust CRISPR/Cas9 System for Convenient, High-Efficiency... into one vector, and plasmids that provide Cas9 variants. All components are then assembled into a single...

  5. 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...

  6. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ... 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...

  7. 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 ...

  8. 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...

  9. 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...

  10. 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...

  11. 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...

  12. 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...

  13. 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...

  14. 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 ...

  15. Modular Cloning Guide

    Type
    Guide
    ...CRISPR/Cas9 vectors expressing multiple gRNAs (up to seven) with a Cas9 nuclease, nickase, dCas9, or dCas9...mechanisms. zCas9i cloning kit: MoClo-compatible CRISPR/Cas9 cloning kit with intronized Cas9 Genome Engineering...assembly of CRISPR/Cas9 constructs for use in plants. The constructs are made with zCas9i, a high efficiency...biology and multigene engineering, including CRISPR/Cas9 components. MoChlo: Modular Cloning Chloroplast ...CRISPR Arnold Driessen 96 plasmids (including CRISPR/Cas9 components) for expression or gene editing in filamentous...constitutive promoters, T7 expression, RBS strength variants, synthetic terminators, protein purification tags...including 11 inducible promoters, 16 RBS strength variants, 8 tag-compatible RBSs, 2 secretion tags, 6 CDSs...

  16. Hot Plasmids: Fall 2025

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    Blog Post
    ...Affordable, low-tech CRISPR/Cas9 experiments By Ashley Waldron CRISPR/Cas9 has revolutionized biotechnology...chromoprotein and gRNA plasmids. Nuclease-dead Cas9 (dCas9) protein is supplied separately. Results can ...concentration in resting neurons. Their soma-targeted variant SomaFRCaMPi also performed substantially better...

  17. 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, ...

  18. 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...

  19. 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...

  20. 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...
Showing: 81 - 100 of 110 results