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TLCV2
(Plasmid #87360)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 87360 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

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

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    Unknown

Gene/Insert

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

Cloning Information

  • Cloning method Unknown
  • 5′ sequencing primer LNCX [5'-AGCTCGTTTAGTGAACCGTCAGATC-3']
  • 3′ sequencing primer EF1a-R [5'CACGGCGACTACTGCACTTA-3']
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Clone sgRNAs into TLCV2 using the same protocol provided with lentiCRISPR v2 (Plasmid #52961, protocol listed under the RESOURCE INFORMATION section). After you have inserted your sgRNA, you should sequence this region with one of the following primers: hU6-F (5'-GAGGGCCTATTTCCCATGATT-3') or LKO.1 5’(5'- GACTATCATATGCTTACCGT-3').

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:

    TLCV2 was a gift from Adam Karpf (Addgene plasmid # 87360 ; http://n2t.net/addgene:87360 ; RRID:Addgene_87360)
  • For your References section:

    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