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Addgene

pLentiGuide
(Plasmid #117986)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 117986 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pCESAx (Addgene 59986)
  • Modifications to backbone
    For sgRNA expression, we modified the sgRNA F+E scaffold (Chen et al., 2013; https://www.addgene.org/59986/ Addgene: U6>sgRNA(F+E) www.addgene.org U6 promoter based on Nishiyama, A. and Fujiwara, S. (2008). RNA interference by expressing short hairpin RNA in the Ciona intestinalis embryo. ) in two ways. First, we exchanged the murine U6 promoter and telomerase-targeting sgRNA with the human U6 promoter, stuffer region, and associated BsmBI cloning sites from lentiCRISPRv2. Additionally, we removed a BsmBI restriction site in the puromycin resistance gene by introducing a non-synonymous mutation.
  • Vector type
    Lentiviral
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    sgRNA backbone (F+E)
  • gRNA/shRNA sequence
    Empty

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Users should follow the oligo cloning protocol for plasmid lentiCRISPR v2 (Addgene #52961) to 1) remove stuffer fragment and 2) insert annealed oligos to complete gRNA.

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:

    pLentiGuide was a gift from Paul Khavari (Addgene plasmid # 117986 ; http://n2t.net/addgene:117986 ; RRID:Addgene_117986)
  • For your References section:

    Coupled Single-Cell CRISPR Screening and Epigenomic Profiling Reveals Causal Gene Regulatory Networks. Rubin AJ, Parker KR, Satpathy AT, Qi Y, Wu B, Ong AJ, Mumbach MR, Ji AL, Kim DS, Cho SW, Zarnegar BJ, Greenleaf WJ, Chang HY, Khavari PA. Cell. 2018 Dec 11. pii: S0092-8674(18)31514-9. doi: 10.1016/j.cell.2018.11.022. 10.1016/j.cell.2018.11.022 PubMed 30580963