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Addgene

pPC1000
(Plasmid #174829)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 174829 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    Lentiviral
  • Vector type
    Mammalian Expression, Lentiviral, CRISPR
  • Selectable markers
    Puromycin ; BFP

Growth in Bacteria

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

Gene/Insert 1

  • Gene/Insert name
    CRISPRi SpCas9 sgRNA cassette
  • Alt name
    EGFP-targeting single guide RNA (modified F+E SpCas9 scaffold) for CRISPRi
  • Species
    Synthetic
  • Insert Size (bp)
    112
  • Mutation
    Detailed in manuscript
  • Promoter mouse U6

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site BstXI (not destroyed)
  • 3′ cloning site BlpI (not destroyed)
  • 5′ sequencing primer mouse U6-forward
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    PuroR-P2A-BFP
  • Alt name
    Puromycin N-acetyl-transferase-P2A-blue fluorescent protein
  • Species
    Synthetic
  • Insert Size (bp)
    1368
  • Promoter EF1a

Cloning Information for Gene/Insert 2

  • Cloning method Ligation Independent Cloning
  • 5′ sequencing primer EF1a-forward
  • 3′ sequencing primer WPRE-reverse
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.
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:

    pPC1000 was a gift from David Liu (Addgene plasmid # 174829 ; http://n2t.net/addgene:174829 ; RRID:Addgene_174829)
  • For your References section:

    Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, Liu DR. Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 10.1016/j.cell.2021.09.018. 10.1016/j.cell.2021.09.018 PubMed 34653350