Human GeCKO library
PurposeA genome-scale CRISPR-Cas9 knockout (GeCKO) library targeting 18,080 genes with 64,751 unique guide sequences enables both negative and positive selection screening in human cells.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Pooled Library||51241||Shipped as suspended DNA in a microcentrifuge tube||1||$500|
This material is available to academics and nonprofits only.
Vector backbonepXRP_001 (lentiCRISPR)
Backbone manufacturerZhang lab
- Backbone size w/o insert (bp) 11590
Vector typeMammalian Expression, Lentiviral, CRISPR
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameHuman GeCKO library
Alt nameTHIS PRODUCT IS DISCONTINUED AS OF 5/8/2014. SEE COMMENTS BELOW FOR DETAILS
- Promoter U6
- Cloning method Restriction Enzyme
- 5′ cloning site BsmBl (not destroyed)
- 3′ cloning site BsmBI (not destroyed)
- 5′ sequencing primer U6 fwd (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
SPECIAL NOTE FROM THE ZHANG LAB: Two new versions of the human GeCKO library have now been released: http://www.addgene.org/crispr/libraries/human_geckov2/ . One version is in the updated lentiCRISPR v2 vector and one is in a 2 plasmid system with Cas9 and sgRNAs on separate plasmids. Both of the updated versions result in higher viral titers, have improved off target score calculations, provide uniform sgRNA coverage, and have better gene and exon coverage. A murine version of the GeCKO library will be available in early summer of 2014.
Human Genome-scale CRISPR Knock-Out Library (GeCKO) as a pool of 65,000 lentiCRISPR constructs with unique sgRNAs (Cas9 single guide RNA). This item is shipped as 1ug of plasmid DNA that will need to be transformed in high-efficiency electrocompetent cells (eg. Lucigen E. Cloni 10G Elite cells) and plated at large scale. Sufficient colonies must be present to keep library representation. Please see included protocol for full details on transformation and plating of bacteria.
Note: Addgene has sequenced part of this library to assure its diversity.
The backbone of this library is lentiCRISPR (http://www.addgene.org/49535/). See map below. The stuffer region downstream of the U6 promoter is replaced by the gRNAs in the library.
In order to provide this library to the scientific community at a reasonable cost, we are shipping this library as DNA in 50 ng/ul (~20ul total) aliquots with blue ice to keep cold.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:Human GeCKO library was a gift from Feng Zhang (Addgene plasmid # 51241)
For your References section:Genome-scale CRISPR-Cas9 knockout screening in human cells. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. Science. 2014 Jan 3;343(6166):84-7. doi: 10.1126/science.1247005. Epub 2013 Dec 12. 10.1126/science.1247005 PubMed 24336571