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

• Depositing Labs: Christopher Vakoc
• Publication: Huang et al. Genes Dev. 2018 Jul 1;32(13-14):915-928. doi: 10.1101/gad.314815.118.

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: ∼20 µL
• Concentration: 50 ng/µL

Resource Information

• Protocols:
  • Library Amplification (117 KB)
  • Library Construction & Screening (158 KB)
• Depositor Data:
  • List of gRNA sequences (330 KB)
  • NGS Read Counts (362 KB)
• Terms and Licenses:
  • UBMTA

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.