Toronto KnockOut (TKO) CRISPR Library - Version 1
(Pooled Library #1000000069)

• Purpose:

  Human CRISPR lentiviral library that has 176,500 guides (12 guides/gene) targeting 17,661 protein-coding genes.

  The library is split into two sub-libraries, each containing 6 sgRNAs per gene. One sub-library (base “90k library”) contains 91,320 sequences and the second sub-library (“supplemental library”) contains 85,180 sequences.

• Vector Backbone:

  pLCKO (Plasmid #73311) - this gRNA backbone does not contain Cas9.

• Depositing Labs: Jason Moffat
• Publication: Hart et al Cell. 2015 Dec 3. 163(6):1515-26. doi: 10.1016/j.cell.2015.11.015.

Library Details

• Species: Human
• Genes targeted: 17,661
• gRNAs: 176,500
  • Base 90k library: 91,320
  • Supplemental library: 85,180
• Controls: Targets include LacZ, EGFP, luciferase, and random loci on chromosome 10
• 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.

Pooled library #1000000069 will be delivered as two tubes, each containing a pooled sub-library. Note that each subpool must be transformed separately.

For each sub-library tube, you will receive:

• Volume: ∼20 µL
• Concentration: 50 ng/µL

Resource Information

• Depositor Data:

  Please visit the Moffat Toronto KnockOut (TKO) library website for the most up-to-date information, including:

  • library sequences and targets
  • read counts
  • RNA-seq data
  
  

  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

One sub-library (base “90k library”) was used in Hart et al (2015) as a first-pass and contains 91,320 sequences. The sgRNAs selected for this library had no genomic off-target sites. Control guides target LacZ, EGFP, luciferase, and random loci on chromosome 10. Additionally, 796 guides identified as promisicuous (20+ predicted binding sites) have been included.

The second sub-library (“supplemental library”) was used in Hart et al (2015) as a second-pass and contains 85,180 sequences. sgRNAs included in this library were selected with the parameter that one potential off-target hit was allowed.

Additionally, the Moffat Lab has deposited the Cas9 and gRNA backbone used in their screen.

• gRNA backbone: pLCKO
• FLAG-tagged Cas9: Lenti-Cas9-2A-Blast

These plasmids are NOT included with library purchase and must be ordered separately.

Figure 1: Moffat Lab Library Logo