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Vakoc Human DNA Binding Domain-Focused CRISPR Knockout Library
(Pooled Library #123334)

  • Purpose
    This lentiviral CRISPR knockout library contains gRNAs that target only the DNA binding domain of human transcription factors. Targeting the functional domain of proteins enhances the severity of the negative selection screen, as reported previously by the Vakoc lab ( Shi et al. Nat Biotechnol 2015).
  • Vector Backbone

    LRG2.1 - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 123334 gRNA pooled library 1 $350 Add to Cart
Available to Academic and Nonprofits Only
  • A Cas9 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with LentiV_Cas9_puro (Addgene #108100) or otherwise with cell lines expressing Cas9.

Library Details

  • Species
    Human
  • DNA binding domains targeted
    1,427
  • gRNAs
    8,658
  • Controls
    36 positive, 50 negative
  • Lentiviral Generation
    3rd

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.

  • Volume
    ∼20µL
  • Concentration
    50ng/µL

Depositor Comments

Lentiviral vectors can recombine during library amplification, resulting in a lower molecular weight band containing the origin and ampicillin resistance cassette. This recombination product should not adversely affect library function as it does not contain lentiviral packaging sequences or the EGFP gene and will not be selected during screening. If a lower molecular weight band is observed after amplification, we recommend starting with a larger prep (gigaprep) from the original sample or gel purifying the amplified library to remove the recombinant band and performing another transformation with the purified sample.

How to cite this pooled library ( Back to top )

These pooled libraries 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:

    Human DNA Binding Domain-Focused CRISPR Knockout library was a gift from Christopher Vakoc (Addgene #123334)
  • For your References section:

    POU2F3 is a master regulator of a tuft cell-like variant of small cell lung cancer. Huang YH, Klingbeil O, He XY, Wu XS, Arun G, Lu B, Somerville TDD, Milazzo JP, Wilkinson JE, Demerdash OE, Spector DL, Egeblad M, Shi J, Vakoc CR. Genes Dev. 2018 Jul 1;32(13-14):915-928. doi: 10.1101/gad.314815.118. PubMed 29945888