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We narrowed to 114 results for: dcas9

Showing: 21 - 40 of 114 results

  2. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without...inhibition, among others. Over the past few years, dCas9 has also become a robust visualization tool to study...University of Massachusetts Medical School developed a dCas9-based multicolor labeling system to visualize chromosomal... al. “Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.” Nature...

  3. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...options include humanized wt Cas9, dCas9 (transcriptional repressor), and dCas9-VP64 (transcriptional activator...D10A nickase mutant (Cas9n), dCas9 (transcriptional repression), and Fok1-dCas9 (dimeric nuclease). Gateway...then digested using BsmBI and ligated into Cas9 or dCas9-containing destination vectors. Destination vector...allows you to assemble type II-A CRISPR arrays for dCas9-based transcriptional repression. The CRISPathBrick...Combinatorial Assembly of Type II-A CRISPR Arrays for dCas9-Mediated Multiplex Transcriptional Repression in...

  4. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...propagation and keeping that dCas9 sequence in the pool. According to this logic, ω-dCas9 variants recognizing... phage (SP) to carry catalytically dead SpCas9 (dCas9) fused to the bacterial polymerase subunit ω. This...NNN PAM sequences adjacent to a protospacer. If ω-dCas9 could recognize a library PAM, it would induce transcription...

  6. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...creating a new dCas9-VPR expression plasmid that uses the piggybac transposase to integrate dCas9-VPR into ... is genomic integration of dCas9-VPR and controlled expression of the dCas9-VPR transgene by doxycycline...methylation by fusing catalytically inactive Cas9 (dCas9) with the catalytic domain from DNA methyltransferase... activator developed by the Church lab, termed “dCas9-VPR”, which facilitates robust activation of target...target genes (20-40x greater than standard dCas9-VP64 activators) when introduced into mammalian cells....activate target genes, the relatively large size of dCas9-VPR limits one’s ability to package it into lentivirus...transfection precluding the need for virus. Furthermore, dCAS9-VPR expression is under the control of the TRE promoter...

  7. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...? Check out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off control all the...repression of protein expression. Addgene has a dCas9 KRAB plasmid for transient knockouts!  Turn it on...

  9. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...capability, and this nuclease-null or “dead” Cas9 (dCas9) variant can then be fused to effector domains such...new cell lines or transgenic animals expressing dCas9 as a means of modulating gene expression (6). Altering... dcas9...

  11. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...strategies for CRISPR-HIV therapeutics - one using dCas9-SAM to activate HIV-1 transcription and destroy ...system available from Addgene. This system uses the dCas9-VP64 fusion commonly used to activate transcription...model. If these escape mutations are frequent, the dCas9-mediated “shock and kill” strategy may be a better...

  13. CRISPR Guide

    Type
    Guide
    ...achieved by using orthogonal dCas9s (e.g. S. pyogenes dCas9 and S. aureus dCas9) tagged with different fluorescent...human cells. The simplest dCas9-based activators and repressors consist of dCas9 fused directly to a single...imaging of proteins in living cells (Figure 9B) dCas9-VPR - dCas9 fused to several different activation domains...utilizes biotin tagging of dCas9 by fusing a biotin acceptor site to dCas9 and co-expressing BirA biotin... defense CRISPRa CRISPR Activation; using a dCas9 or dCas9-activator with a gRNA to increase transcription...target gene CRISPRi CRISPR Interference; using a dCas9 or dCas9-repressor with a gRNA to repress/decrease transcription...one strand. Catalytically inactive “dead” Cas9 (dCas9) contains mutations in both nuclease domains, D10A...

  14. Plan Your Experiment

    Type
    Guide
    ...permanently modifying the genome dCas9-repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting promoter elements... elements of target gene dCas9-KRAB is more effective than dCas9 alone for mammalian cell lines. Activate...without permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter elements...activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs should be targeted to...

  15. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ... 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...histone tail (H3K4me0) to a DNA-binding protein (dCas9 shown here). D3L recruits the endogenous, autoinhibited...

  16. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...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...background labeling. By fusing miniTurboID with dCas9 to create TurboCas, the Shilatifard Lab created ...

  17. The 12 Days of CRISPR

    Type
    Blog Post
    ...🎵 On the 3rd day of CRISPR, Addgene gave to me: dCas9-based gene editing for cleavage-free genomic knock-in...

  18. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ... editor targets DNA (via catalytically inactive dCas9) to modify chromatin. A fluorescent protein can ...Jacobs-Wagner’s lab produce different levels of dCas9 expression and contain different antibiotic resistance...

  19. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...antibiotic resistance selection markers, gRNAs, and dCas9 proteins. Second, it can be introduced into a broader...promoters, selection markers, and different versions of dCas9. Following integration, Mobile-CRISPRi is stably...

  20. CRISPR Cheat Sheet

    Type
    Blog Post
    ...versions of CRISPR nucleases that can’t cut DNA (dCas9, for example) that are used to increase or decrease...
Showing: 21 - 40 of 114 results