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pET-dCas9-VP64-6xHis
(Plasmid #62935)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 62935 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET29
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5161
  • Total vector size (bp) 9544
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Mach1
  • Growth instructions
    For protein expression, transform into BL21 Star (DE3) cells. Induce at 0.6 OD with 0.5 mM IPTG final concentration. Growing overnight (16 hours) at 20-25 C.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    dCas9-VP64
  • Species
    S. pyogenes
  • Insert Size (bp)
    4383
  • Mutation
    D10A and H840A amino acid changes render Cas9 nuclease inactive for DNA cleavage
  • Promoter T7
  • Tags / Fusion Proteins
    • 3xFLAG (C terminal on insert)
    • NLS (C terminal on insert)
    • VP64 (C terminal on insert)
    • 6xHis (C terminal on backbone)

Cloning Information

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    pMJ806 served as template for Cas9 gene. This was reported in Pattanayak et. al. Nature Biotechnology 2013.
  • Terms and Licenses
  • Article Citing this Plasmid

Depositor Comments

Alternative plasmid name: pJAZ0007

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pET-dCas9-VP64-6xHis was a gift from David Liu (Addgene plasmid # 62935 ; http://n2t.net/addgene:62935 ; RRID:Addgene_62935)
  • For your References section:

    Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Zuris JA, Thompson DB, Shu Y, Guilinger JP, Bessen JL, Hu JH, Maeder ML, Joung JK, Chen ZY, Liu DR. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30. 10.1038/nbt.3081 PubMed 25357182