Vakoc Human Kinase Domain-Focused CRISPR Knockout Library
(Pooled Library #117725)

• Purpose: This lentiviral CRISPR knockout library contains gRNAs that target only the kinase domain of protein kinases. 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 (Plasmid #108098) - does not express Cas9

• Depositing Labs: Christopher Vakoc
• Publication: Tarumoto et al. Mol Cell. 2018 Mar 15;69(6):1017-1027.e6. doi: 10.1016/j.molcel.2018.02.011.

Library Details

• Species: Human
• Genes targeted: 482 kinase genes (496 domains)
• gRNAs: 3,051
• Controls: 36 positive, 51 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 and Sequencing Protocol (DOCX, 157 KB)
• Depositor Data:
  • List of gRNA sequences (XLSX, 133 KB)
  • NGS Read Counts (TXT, 87 KB)
• Articles Citing this Pooled Library:
  • Human Kinase Domain-Focused CRISPR KO (#117725)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

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.