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lentiCRISPR v2-dCas9
(Plasmid #112233)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 112233 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    Custom
  • Backbone size w/o insert (bp) 10000
  • Total vector size (bp) 14873
  • Vector type
    Mammalian Expression, Lentiviral, CRISPR
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    30°C
  • Growth Strain(s)
    NEB Stable
  • Growth instructions
    Only amplify in RecA- bacteria
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    dCas9
  • Species
    Synthetic
  • Insert Size (bp)
    4200
  • Mutation
    Changed: Aspartic acid 10 to Glycine, Histidine 840 to Alanine
  • Promoter EFS-NS
  • Tag / Fusion Protein
    • FLAG (C terminal on insert)

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site AgeI, XbaI, AfeI (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer hEF1aprom-F
  • 3′ sequencing primer WPRE-R
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    Porumycin resistance
  • Alt name
    puromycin N-acetyl-transferase
  • Alt name
    PAC
  • Insert Size (bp)
    600
  • Promoter EFS-NS

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site BanHI (not destroyed)
  • 3′ cloning site MluI (not destroyed)
  • 5′ sequencing primer Puro-F2
  • 3′ sequencing primer WPRE-R
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    addgene
  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry

Depositor Comments

Plasmid is a derivative of Addgene Plasmid #52961, where the Cas9 gene has been mutated at codons 10 and 840 to create a catalytically inactive version of Cas9 (dCas9).

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:

    lentiCRISPR v2-dCas9 was a gift from Thomas Gilmore (Addgene plasmid # 112233 ; http://n2t.net/addgene:112233 ; RRID:Addgene_112233)
  • For your References section:

    CRISPR/Cas9-based editing of a sensitive transcriptional regulatory element to achieve cell type-specific knockdown of the NEMO scaffold protein. Babaei M, Liu Y, Wuerzberger-Davis SM, McCaslin EZ, DiRusso CJ, Yeo AT, Kagermazova L, Miyamoto S, Gilmore TD. PLoS One. 2019 Sep 25;14(9):e0222588. doi: 10.1371/journal.pone.0222588. eCollection 2019. 10.1371/journal.pone.0222588 PubMed 31553754