Human GeCKO library
(Pooled library #51241)

• Purpose: A 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.
• Depositing Lab: Feng Zhang
• Publication: Shalem et al Science. 2014 Jan 3;343(6166):84-7. doi: 10.1126/science.1247005. Epub 2013 Dec 12.

Backbone

• Vector backbone: pXRP_001 (lentiCRISPR)
• Backbone manufacturer: Zhang lab
• Backbone size w/o insert (bp): 11590
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Human GeCKO library
• Alt name: THIS PRODUCT IS DISCONTINUED AS OF 5/8/2014. SEE COMMENTS BELOW FOR DETAILS
• Species: Synthetic
• Promoter: U6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsmBl (not destroyed)
• 3′ cloning site: BsmBI (not destroyed)
• 5′ sequencing primer: U6 fwd

Resource Information

• Supplemental Documents:
  • GeCKO Libary Transformation Protocol
  • List of genes and sgRNA sequences in library

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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.
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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.