Toronto KnockOut (TKO) CRISPR Library - Version 1
(Pooled Library #1000000069)
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 BackbonepLCKO- this gRNA backbone does not contain Cas9
- Depositing Labs
|Item||Catalog #||Description||Quantity||Price (USD)|
|Pooled Library||1000000069||Two gRNA pooled half-libraries †||1||$500||Add to Cart|
† A Cas9 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with Lenti-Cas9-2A-Blast (Addgene#73310) or otherwise with cell lines already expressing Cas9.
- Base 90k library: 91,320
- Supplemental library: 85,180
ControlsTargets include LacZ, EGFP, luciferase, and random loci on chromosome 10
This library will be delivered as two pooled DNA half-libraries in microcentrifuge tubes on blue ice. The tubes' contents will not necessarily be frozen. For best results, minimize freeze-thaws.
For each half-library, you will receive:
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.
Note that each subpool must be transformed separately.
Terms and Licenses
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.
These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the pooled libraries were described, and include Addgene in the Materials and Methods of your future publications.
Example for your Materials & Methods section:Toronto human knockout pooled library (TKO) was a gift from Jason Moffat (Addgene #1000000069)
For your References section:High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities . Hart T, Chandrashekhar M, Aregger M, Steinhart Z, Brown KR, MacLeod G, Mis M, Zimmermann M, Fradet-Turcotte A, Sun S, Mero P, Dirks P, Sidhu S, Roth FP, Rissland OS, Durocher D, Angers S, Moffat J. Cell. 2015 Dec 3;163(6):1515-26. 10.1016/j.cell.2015.11.015.PubMed 26627737