Garnett Lab MinLibCas9 Library
(Pooled Library #164896)

• Purpose:

  This human genome-wide CRISPR knockout library is designed with two single guide RNAs (sgRNA) per gene.

  The minimal library has been optimized through empirical assessment using comprehensive sgRNA efficiency metrics.

• Vector Backbone:

  pKLV2-U6gRNA(BbsI)-ccdB-PGKpuro2ABFP-W - does not express Cas9

• Depositing Labs: Mathew Garnett
• Publication: Goncalves et al Genome Biol. 2021 Jan 21;22(1):40. doi: 10.1186/s13059-021-02268-4.

Library Details

• Species: Human
• Genes targeted: 18,761
• gRNAs: 37,722
• Controls: 200 non-targeting sgRNAs
• Lentiviral Generation: 3rd

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: ∼20 µL
• Concentration: 50 ng/µL

Resource Information

• Protocols:
  • Propagation of MinLibCas9 Library (DOC, 88 KB)
  • Primers and PCR protocols to prepare samples of amplified library for deep sequencing verification (DOC, 133 KB)
• Depositor Data:
  • Table of target genes and gRNA sequences (XLS, 7.2 MB)
  • gRNA Read Counts (XLS, 2.2 MB)
• Terms and Licenses:
  • UBMTA
  • Evrogen Limited Use Label License for FPs

Depositor Comments

• Garnett Lab MinLibCas9 Library.
  Figure 2. Goncalves et al. Genome Biol. 2021

The depositing lab recommends calculating total read counts per gRNA using the count_spacers script from the Feng Zhang Lab.

Read distribution per gRNA can be determined using the ineq package in R, calculating both the Lorenz-curve and Gini-coefficient.