Pooled Libraries
Toronto KnockOut (TKO) CRISPR Library v3

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

Available in either a 1 vector (lentiCRISPRv2 backbone) or 2 vector (pLCKO2 backbone) system.

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. (How to cite )

Ordering

Item	Catalog #	Description	Quantity	Price (USD)
Pooled Library	90294	gRNA pooled library in lentiCRISPRv2 Original TKOv3	1		$500	Add to Cart
Pooled Library	125517	gRNA pooled library in pLCKO2	1		$500	Add to Cart

Available to Academic and Nonprofits Only

Library Details

Species
Human
Genes targeted
18,053
gRNAs
70,948
Controls
142 guides targeting LacZ, EGFP, and luciferase
Lentiviral Generation
3rd

Library Shipping

This library will be delivered as a pooled DNA library 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
- CRISPR Screen Protocol (244 KB)
Depositor Data
- TKOv3 Guide Sequences (3.4 MB)
Please also visit the Moffat Toronto KnockOut (TKO) library website for the most up-to-date information.

The Moffat Lab also provides a gRNA viewer, that displays gRNA sequence information for a specific gene.
Terms and Licenses
- UBMTA

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 4 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 without Cas9, that can be utilized in cells that stably express Cas9.

Moffat Lab Library Logo

How to cite this pooled library ( Back to top )

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 (TKOv3) was a gift from Jason Moffat (Addgene #)
For your References section:

For #90294: Evaluation and Design of Genome-Wide CRISPR/SpCas9 Knockout Screens. Hart T, Tong AHY, Chan K, Van Leeuwen J, Seetharaman A, Aregger M, Chandrashekhar M, Hustedt N, Seth S, Noonan A, Habsid A, Sizova O, Nedyalkova L, Climie R, Tworzyanski L, Lawson K, Sartori MA, Alibeh S, Tieu D, Masud S, Mero P, Weiss A, Brown KR, Usaj M, Billmann M, Rahman M, Constanzo M, Myers CL, Andrews BJ, Boone C, Durocher D, Moffat J. G3 (Bethesda). Aug 7;7(8):2719-2727. doi: 10.1534/g3.117.041277. PubMed 28655737

For #125517: Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies. Mair B, Tomic J, Masud SN, Tonge P, Weiss A, Usaj M, Tong AHY, Kwan JJ, Brown KR, Titus E, Atkins M, Chan KSK, Munsie L, Habsid A, Han H, Kennedy M, Cohen B, Keller G, Moffat J. Cell Rep. 2019 Apr 9;27(2):599-615.e12. doi: 10.1016/j.celrep.2019.02.041. PubMed 30970261

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