Mouse CRISPR Proximal Poly(A) Site Knockout Library
(Pooled Library #190744)

• Purpose:

  The mouse pKO library targets proximal poly(A) signal sequences in genes identified to be differentially polyadenylated in the mouse B16-F10 melanoma system. Growth controls include gene knockouts of core essential or tumor suppressor genes and a large number of control paired-guide RNAs targeting poly(A) sites in genes that are not expressed in B16-F10 melanoma cells. Deleting the proximal poly(A) site forces use of more distal sites.

• Vector Backbone:

  lentiGuide-Puro - does not express Cas9

• Depositing Labs: Robert Bradley
• Publication: Gabel et al. Nat Commun. 2024 Feb 1;15(1):959. doi: 10.1038/s41467-024-44931-x.

Library Details

• Species: Mouse
• Genes targeted: 144 proximal poly(A) sites targeted
• gRNAs: 8-10 pgRNAs per target
• Controls:
  • 150 control pgRNAs targeting poly(A) sites in genes that are not expressed in B16-F10 cells
  • 150 growth control pgRNAs targeting exonic regions in 15 core essential genes, tumor suppressor genes, or immune system relevant genes
• Total number of gRNAs: 1718 pgRNAs
• 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:
  • The depositor recommends following the GeCKo v2.0 Library Amplification Protocol (PDF, 272 KB)
  • Paired gRNA NGS Protocol (PDF, 234 KB)
• Depositor Data:
  • List of pgRNA sequences (XLS, 540 KB)
  • pgRNA NGS Read Counts (TXT, 99 KB)
  • Histogram of reads detected per pgRNA (PDF, 5 KB)
  • Density plot of reads detected per pgRNA (PDF, 9 KB)
• Terms and Licenses:
  • UBMTA