• PURPOSE

  LentiCRISPR v2 was modified into an all-in-one dox inducible system. The addition of doxycycline induces Cas9-2A-eGFP. The U6 promoter drives constitutive sgRNA expression.

BACKBONE

• Vector backbone
  lentiCRISPR v2
• Backbone manufacturer
  Feng Zhang lab
• Backbone size w/o insert (bp)16733
• Modifications to backbone
  LentiCRISPR v2 was modified into an all-in-one inducible system, doxycycline induces Cas9-P2A-GFP expression. sgRNA and puromycin expression are driven by constitutive promoters. The following modifications were made to lentiCRISPR v2: the EF1 promoter was removed and replaced with a tight TRE promoter; P2A-puromcycin was removed and replaced with T2A-eGFP; and a EF1 promoter-puromycin-P2A-rtTA cassette was added.
• Vector type
  Mammalian Expression, Lentiviral, CRISPR ; Doxycycline inducible; eGFP reporter
• Selectable markers
  Puromycin

GENE/INSERT

• Gene/Insert name
  Cas9-2A-eGFP
• PromoterTight TRE promoter

Pan-Cancer Analyses Reveal Genomic Features of FOXM1 Overexpression in Cancer. Barger CJ, Branick C, Chee L, Karpf AR. Cancers (Basel). 2019 Feb 21;11(2). pii: cancers11020251. doi: 10.3390/cancers11020251. 10.3390/cancers11020251 PubMed 30795624