CRISPR Human Genome Knockout Libraries

Cellecta’s latest Single-Module Human Genome-Wide CRISPR Knockout Libraries are designed to deliver robust knockout performance, enabling efficient pooled screening across the human genome. 

• • Single-module, pooled sgRNA libraries targeting approximately 19,382 protein-coding genes and 1,859 microRNAs 

• • • 87K Human CRISPR Knockout Library (4 sgRNAs/gene) comprises ~87,000 sgRNA constructs. In addition to gene-targeting guides, it includes 1,000 non-targeting controls, 400 intron-targeting controls, 400 intergenic sgRNAs, and 400 safe-harbor sgRNAs targeting 2 sites: AAVS1 and CLYBL.
    • 43K Human CRISPR Knockout Library (2 sgRNAs/gene)comprises ~43,000 sgRNA constructs. In addition, it includes 200 non-targeting controls and 200 intergenic controls, and achieves the same knockout efficiency as the 87K library despite its more compact design.

• • Both the 43K and 87K libraries are designed using the criteria for effective guides described by DeWeirdt et al. 2022, which generates more robust screening results than the standard knockout 80K v2 library, assessed by the observed depletion of sgRNAs targeting common essential genes (Fig. 1)

Fig 1: Cellecta Next-Generation Human CRISPR 43K and 87K Knockout Libraries Outperform CRISPR 80Kv2 Knockout Library. The 87K design offers 4 sgRNAs/gene, while the 43K has a compact 2 sgRNAs/gene design. The reduced library size enables effective screens with fewer cells, as it achieves knockout efficiencies comparable to those of the 87K Knockout library.  

• • Guide designs are obtained using the CRISPick web tool from the Genetic Perturbation Platform (GPP) at the Broad Institute (PMID: 36068235) and are then ranked using Cellecta's proprietary algorithm.
    
  • The tracrRNA portion of the sgRNA features the HECG design, an improvement over the HEAT design. 
    
  • These libraries may be obtained in the following vectors:
    • UbiC-TagRFP and Puro (pRSG162-U6-sg-UbiC-TagRFP-2A-Puro vector)
      
    • UbiC-Puro (pRSGUPS2-U6-sg-UbiC-Puro vector)
      

• • Plasmid and lentiviral packaged formats 
    • 200 ug plasmid
    • 2 x 10^8 TU or 1 x 10^9 TU

*NOTE: Researchers who would like to purchase more units of our older libraries may contact us at info@cellecta.com

Manual and Product Documentation

• Brochure: Cellecta CRISPR & RNAi Functional Genomics Capabilities
• Gene List: 
  • CRISPR Human Genome 87K Knockout Library Gene List
  • CRISPR Human Genome 43K Knockout Library Gene List
• Manual: CRISPR Pooled Lentiviral sgRNA Libraries Manual

Library Vector Information

* Sequences are available in FASTA (.txt), GenBank (.gb), and SnapGene (.dna) formats. You may use the free SnapGene Viewer to open .dna files. If you would like vector files in another format, please contact us. To download .txt and .gb files, right click on the link and Save As. 

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Alignment Software Supporting Files

• Library Configuration Files (JSON format, compressed): 
  • KOHGW-87k Libraries
  • KOHGW-43k Libraries

• Sample Description Input Form: XLSM (Please enable macros)