Mouse Kinome CRISPR Knockout Library (Brie)
(Pooled Libraries #75316, #75317)

• Purpose:

  Mouse kinome CRISPR knockout pooled libraries. Each library contains 2,852 unique sgRNAs targeting 713 mouse kinase genes with four guides per target.

• Vector Backbone:

  • lentiCRISPR v2 (Plasmid #52961) backbone (one-vector system) - expresses Cas9
  • lentiGuide-Puro (Plasmid #52963) backbone (two-vector system) - does not express Cas9

• Depositing Labs: John Doench, David Root
• Publication: Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.

Library Details

• Species: Mouse
• Genes targeted: 713
• gRNAs: 2,852
• Controls: 100
• Lentiviral Generation: 3rd

Library Shipping

Each library is delivered in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

• Volume: ∼10 µL
• Concentration: 10 ng/µL

Resource Information

• Protocols:
  • Library Amplification Protocol (PDF, 90 KB)
  • Library Sequencing Protocol (PDF, 162 KB)
• Depositor Data:
  • Mouse Kinome Library Target Genes (ZIP, 313 KB)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Addgene Notes

The amplification protocol provided, which recommends using 400 ng of DNA per transformation, was written in reference to the whole genome-wide libraries. These kinome-specific sub-pools contain fewer plasmids and thus require less DNA to achieve complete representation. Therefore, using the 100 ng of DNA as provided for transformation is sufficient.

NOTE: Lentiviral vectors can recombine during library amplification, resulting in a band of ~1 kb 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 PuroR gene and will not be selected during screening. If recombination 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.