URL: http://www.addgene.org/59702
Proper Citation: RRID:Addgene_59702
Insert Name: EGFP sgRNA
Organism: Synthetic
Bacterial Resistance: Ampicillin
Defining Citation: PMID:25184501
Vector Backbone Description: Vector Backbone:pLKO.1; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
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.
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Source: Addgene