Human CRISPR Knockout Library (H3)
(Pooled library #133914)

• Purpose:

  The H3 genome-wide human CRISPR KO library targets more than 18,000 annotated genes in the human genome with 6 gRNAs per gene on average. gRNAs are 19 nucleotides in length and are selected based on computationally-predicted and experimentally-observed maximum KO efficiency. Users can conduct genome-wide CRISPR/Cas9-mediated KO screens in human cells with this library.

• Vector Backbone:

  sgRNA_2.0 - does express Cas9 (map)

• Depositing Labs: Xiaole Shirley Liu, Myles Brown
• Publication: Unpublished (How to cite )

Library Details

• Species: Human
• Genes targeted: >18,000
• gRNAs: 117,587
• Lentiviral Generation: 3rd
• Controls: 3,842 targeting AAVS1 (3,540 gRNAs), ROSA26 (203 gRNAs) and CCR5 (99 gRNAs)

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

• Volume: ∼40 µL
• Concentration: 25 ng/µL

Resource Information

• Protocols:
  • Library Amplification and NGS Protocol (DOCX, 38 KB)
  • Screen Protocol — Adherent Cells (DOCX, 279 KB)
  • Screen Protocol — Suspension Cells (DOCX, 280 KB)
• Depositor Data:
  • H3 gRNA Sequences (TXT, 4.4 MB)
  • H3 gRNA Distribution Plot (PNG, 124 KB)
• Articles Citing this Pooled Library:
  • Human CRISPR KO (H3) (#133914)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  Industry Terms:
  • Not Available to Industry
  Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

In designing the library, the 6 top-performing gRNAs were selected for each gene based on computational prediction and experimental evidence on KO efficiency of individual gRNAs in the Brunello library (the Broad Institute, Sanson et al., Nat Comm, 2018), our lab’s previous design of genome-wide CRISPR KO libraries ( Xu et al. Genome Res, 2015; Chen et al. Bioinformatics, 2018) and relevant publications in the field ( Schoonenberg et al., Genome Biol, 2018).

The depositing lab recommends the MAGeCK pipelines for computational analysis of screens conducted using this library: MAGeCK and MAGeCKFlute.

Please note that although there are 117,587 total gRNAs in the plasmid library, 647 "negative control" gRNAs have been removed from the list of gRNA sequences. This is because, in practice, these gRNAs, which were originally designed as negative control gRNAs, have significant unintended enrichment or depletion in cellular screens and are thus not suitable for control purposes. They have therefore been removed from the sequence files to avoid being referenced in downstream computational analysis.