Toronto KnockOut (TKO) CRISPR Library - Version 3
(Pooled Libraries #90294, #125517)

• Purpose:

  Human CRISPR lentiviral library that has 70,948 guides (4 guides/gene) targeting 18,053 protein-coding genes.

• Vector Backbone:

  • lentiCRISPR v2 (Plasmid #52961) backbone (one-vector system) - expresses Cas9
  • pLCKO2 (Plasmid #125518) backbone (two-vector system) - does not express Cas9

• Depositing Labs: Jason Moffat
• Publication: Hart et al G3 (Bethesda). 2017 Aug 7;7(8):2719-2727. doi: 10.1534/g3.117.041277.
  
  Mair et al Cell Rep. 2019 Apr 9;27(2):599-615.e12. doi: 10.1016/j.celrep.2019.02.041.

Library Details

• Species: Human
• Genes targeted: 18,053
• gRNAs: 70,948
• Controls: 142 guides targeting LacZ, EGFP, and luciferase
• 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: ∼20 µL
• Concentration: 50 ng/µL

Resource Information

• Protocols:
  • Library Screening Protocol (PDF, 250 KB)
• Depositor Data:
  • TKOv3 Guide Sequences (XLSX, 3.5 MB)

  The Moffat Lab also provides a gRNA viewer , that displays gRNA sequence information for a specific gene.

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

The Toronto KnockOut Library v3 (TKOv3) contains 70,948 gRNAs targeting 18,053 protein coding genes (4 gRNAs/gene) with 142 control gRNAs targeting EGFP, LacZ, and luciferase (71,090 total).

The gRNAs in this library are optimized for editing efficiency using empirical data described in the associated article. As a result, the number of gRNAs per gene is decreased to four based on the finding that there are diminishing benefits to more gRNAs per gene beyond four. The optimized library size of TKOv3 offers the benefit of improved accuracy, efficiency, and scalability for CRISPR screens.

This library is available in two formats: 1) as a one-component library that contains Cas9, alleviating the need to include a separate Cas9 and 2) as a two-component system without Cas9, that can be utilized in cells that stably express Cas9.