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pXPR_011
(Plasmid #59702)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 59702 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pLKO.1
  • Total vector size (bp) 8395
  • Vector type
    Mammalian Expression, Lentiviral, CRISPR
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Stbl3
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    EGFP sgRNA
  • Species
    Synthetic
  • Promoter hU6

Cloning Information

Resource Information

Depositor Comments

This lentiviral vector can be used to assay Cas9 activity. In the absence of Cas9, the cells are both resistant to puromycin and express EGFP. In the presence of functional levels of Cas9, the sgRNA expressed from the U6 cassette targets EGFP, and thus cells will still be puromycin-resistant, but will no longer express EGFP. The fraction of EGFP-positive and -negative cells can be accurately quantitated by flow cytometry. Please note that it is unlikely that 100% of cells will ever be EGFP-negative, as a fraction of cells are likely to have in-frame indels, and those the cells may still be green. This assay is best done with cells infected with pXPR_011 at an MOI <1.

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:

    pXPR_011 was a gift from John Doench & David Root (Addgene plasmid # 59702 ; http://n2t.net/addgene:59702 ; RRID:Addgene_59702)
  • For your References section:

    Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation. Doench JG, Hartenian E, Graham DB, Tothova Z, Hegde M, Smith I, Sullender M, Ebert BL, Xavier RJ, Root DE. Nat Biotechnol. 2014 Sep 3. doi: 10.1038/nbt.3026. 10.1038/nbt.3026 PubMed 25184501