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We narrowed to 21 results for: dcas9 vp64

Showing: 1 - 21 of 21 results

  1. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ... lines.   dCas9-VP64 Description CRISPR activation can occur through fusing dCas9 with VP64, a strong...shown significant improvements upon the initial dCas9-VP64 method, so there are multiple options to choose...transcriptional activation domain. Guided by dCas9, VP64 recruits transcriptional machinery to specific... on which sequence is targeted. Performance dCas9-VP64 activation is generally thought of as the “first... been able to achieve much higher activation, dCas9-VP64 is great for experiments that requires modest...transcription. This is done through creating a dCas9/VP64 fusion protein engineered with aptamers that ...Description Rather than using a single copy of VP64 per each dCas9, SunTag uses a repeating peptide array to...

  2. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...genes using dCas9-based activators The simplest of the dCas9 activators consists of dCas9 fused to a single...Repressing target genes using dCas9-based repressors Early experiments using dCas9 in bacteria demonstrated...generally the strongest activator, consists of a dCas9-VP64 fusion and a modified gRNA that is capable of... Fig. 3: Cas9 repressors include dCas9 alone and a dCas9-KRAB fusion, both targeted to the promoter...region. Cas9 activators include Cas9 fused to VP64, VP64 along with other SAM factors, and the Suntag ...even making a break.  Base editors are dead Cas9 (dCas9) or nCas9 enzymes fused to either cytosine or adenosine...genome but still want to affect gene expression? dCas9 can be used as a platform to deliver various cargo...

  3. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...specific genes using dCas9-VP64 activator fragments (dCas9(C)-FKBP-2xNLS-VP64 and dCas9(N)-FRB-NES). This...gene activation when fused to dCas9. In the dCas9-SunTag-VP64 system, dCas9 is fused to a scaffold containing...domains onto dCas9 to generate a dCas9-VP64-p65-Rta fusion protein (referred to as dCas9-VPR) had a synergistic...greater than the original dCas9-VP64 activator. They also revealed that dCas9-VPR mediated gene activation...proven challenging using the existing dCas9-VP64 activator. The dCas9-VPR activation plasmids have great ...on a Gaussia luciferase reporter vector. dCas9 or dCas9-VP64 (a control that is known to enhance gene ...62883 &62884; 62885 & 62886) Rapamycin-inducible dCas9-VP64 activator set (Addgene plasmids 62887 & 62888...

  4. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...transcriptional activator domain (VP64 or p65). Upon expression in the cell, the dCas9-CIB1 fusion binds to the...fused to both the N- and C-termini of dCas9; and 2) CRY2FL-VP64, a fusion of full-length CRY2 and the ...in which dCas9 binding occurs. Bubeck et al. expressed a telomere-targeting gRNA and a dCas9 fusion protein.... sphaeroides Cas9, dCas9 Gene editing Zhou et al., 2018   pdDronpa Cas9, dCas9 Gene editing, transcription...Shining light on transcriptional activation using dCas9 Initial photoactivatable CRISPR systems published...genomic anchor - an inactive, dead Cas9 protein (dCas9) fused to CIB1; and 2) the activator - the CRY2 ...proteins. The best performing combination was NLS-dCas9-trCIB1 and NLSx3-CRYPHR-p65 - it had the lowest ...

  5. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...including pcDNA-dCas9-p300 Core and pcDNA3.3-Nm-dCas9-p300 Core, as well as pLV-dCas9-p300-P2A-PuroR for...pCMV-dCas9-D3A) and Tet-dependent (TetO-dCas9-D3A) constructs. For lentiviral expression, Fuw-dCas9-Dnmt3a...Fusing dCas9 to a transcriptional activator like VP64 or VPR activates transcription, whereas dCas9-KRAB...activators/repressors - dCas9-p300 increased transcriptional activation more than dCas9-VP64, especially when...Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase...Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety of epigenetic modifiers to specific...Transcriptional activation p300 acetyltransferase dCas9 fused to the catalytic domain of p300 acetyltransferase...

  7. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...inactive form of Cas9 known as dCas9 (Jinek et al., 2012). Like Cas9, dCas9 can bind to a specific DNA sequence...targeting gRNA. But dCas9 does not cleave the DNA. Much of the research using dCas9 has focused on transcriptional...mammalian cells by expressing dCas9 with an N-terminal 3xFLAG tag (3xFLAG-dCas9) and a gRNA targeting their... a fusion to a transcriptional activator such as VP64 (Gilbert et al., 2013), or repression of transcription...activators (Qi et al., 2013). However, the fusion of dCas9 with a protein tag allows for the isolation of a...genomic region using the CRISPR system, consisting of dCas9 and targeting gRNA, to identify molecular interactions...3xFLAG tag, allowed these researchers to isolate dCas9 with the bound genomic locus, which they then confirmed...

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

    Type
    Blog Post
    ...an inactive RNA-guided DNA binding protein (dCas9). dCas9 is then tethered to a repressive domain, such...loss of function while nuclease-deactivated Cas9 (dCas9) can be tethered to a repressor (CRISPRi) or an ...library In addition to inhibiting gene expression, dCas9 can also be used for transcriptional activation....developed to recruit transcriptional machinery to dCas9 and its RNA directed binding site. These methods... fusion of an activation domain such as VP16 to dCas9 or the recruitment of multiple copies of VP16 termed... with only two constructs (the other containing dCas9-VP16). Like CRISPRi, CRISPRa is only effective when... activator design that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA aptamers, and the  MS2...

  9. Lentiviral Prep Service

    Type
    Collection
    ...Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP... dCAS-VP64_Blast dCAS9 (D10A, N863A) Blasticidin 3rd generation lenti vector encoding dCAS9-VP64 with ...with 2A Blast resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE) Zhang Pooled Barcoding Libraries...56,762 unique sgRNAs. This backbone does not contain dCas9. 56,762 unique sgRNAs targeting 18,885 human genes...Human CRISPR inhibition library Dolcetto in XPR_050 (VP64) Set A Human CRISPR inhibition library in backbone...non-targeting controls. This backbone does not contain dCas9. 57,050 unique sgRNAs targeting 18,901 human genes...Human CRISPR inhibition library Dolcetto in XPR_050 (VP64) Set B Human CRISPR inhibition library in backbone...

  11. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...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...

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

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

  13. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...target genes (20-40x greater than standard dCas9-VP64 activators) when introduced into mammalian cells....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...Gersbach pcDNA3.1-GoldenGate, pcDNA3.1-GoldenGate-VP64 Maria-Elena Torres-Padilla pTALYM3, pTALYM4 ... activator developed by the Church lab, termed “dCas9-VPR”, which facilitates robust activation of target...activate target genes, the relatively large size of dCas9-VPR limits one’s ability to package it into lentivirus...

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

    Type
    Blog Post
    ...tools. CRISPRi uses a nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily block the transcription...hand, recruits transcriptional activators such as VP64, SAM, or SunTag to specific ncRNA loci to enhance...

  15. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...environmental triggers. This is done by creating a split dCas9 protein that can reassemble when exposed to specific...Cas9 activators, check out our blog post covering VP64, Suntag, SAM, and VPR. Worry less about off targets...

  16. Zhang Lab CRISPR Page

    Type
    Collection
    ...for transient transfection : 61422 : Expresses dCAS9-VP64 activator with 2A GFP 61423 : Expresses the MS2...production : 61425 : lentiviral vector encoding dCAS9-VP64 with 2A Blast resistance marker 61426 : lentiviral...three components: A nucleolytically inactive Cas9-VP64 fusion A sgRNA incorporating two MS2 RNA aptamers...

  17. Plan Your Experiment

    Type
    Guide
    ...genome sequence dCas9 or dCas9 fusion with transcriptional repressor (such as dCas9-KRAB) dCas9-KRAB can be...the genome sequence dCas9 fusion with transcriptional activator (such as dCas9-VP64) Many different activator...edits without double-strand breaks Base editor (dCas9 or Cas9 nickase fusions with different base editing...be more effective than dCas9 alone for mammalian cell lines Interfere Activate (CRISPRa) Increase gene... the transcription start site. CRISPRa requires dCas9 to be fused to transcriptional activators that need...N. J. (2023). Elongation roadblocks mediated by dCas9 across human genes modulate transcription and nascent...

  18. 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...(Figure 9C) SAM activators - co-expression of dCas9-VP64 with a modified scaffold gRNA and additional ...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...

  19. Plan Your Experiment

    Type
    Collection
    ...genome sequence dCas9 or dCas9 fusion with transcriptional repressor (such as dCas9-KRAB) dCas9-KRAB can be...the genome sequence dCas9 fusion with transcriptional activator (such as dCas9-VP64) Many different activator...edits without double-strand breaks Base editor (dCas9 or Cas9 nickase fusions with different base editing...be more effective than dCas9 alone for mammalian cell lines Interfere Activate (CRISPRa) Increase gene... the transcription start site. CRISPRa requires dCas9 to be fused to transcriptional activators that need...N. J. (2023). Elongation roadblocks mediated by dCas9 across human genes modulate transcription and nascent...

  20. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ...Mammalian Lentiviral expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent...separate accessory pack is available for dCas9 and FokI-dCas9. Yamamoto Multiplex CRISPR/Cas9-based genome...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...Mammalian U6 yes, nick S. pyogenes Conklin pXFokI-dCas9 60901 Mammalian U6 yes, FokI S. pyogenes Conklin...Mammalian U6 none S. aureus Joung pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-Puro 71236 Mammalian/Lentiviral hU6 yes...Lentiviral U6 none S. pyogenes Puro Zhang pRS416-dCas9-Mxi1 + TetR + pRPR1(TetO)-NotI-gRNA 73796 Yeast ...pyogenes Chen pdCas9-DNMT3A-EGFP 71666 Mammalian U6 yes, methylation S. pyogenes EGFP Zoldos pdCas9-DNMT3A-PuroR...

  21. CRISPR Guide

    Type
    Collection
    ...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...(Figure 9C) SAM activators - co-expression of dCas9-VP64 with a modified scaffold gRNA and additional ...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...
Showing: 1 - 21 of 21 results