MYC-CRISPR Library
(Pooled Library #173195)

• Purpose:

  This pooled lentiviral CRISPR knockout library targets E-boxes genome-wide. It was designed based on MYC-ChIP-seq data from several MYC-dependent cancer cell lines. It can be applied for identification of essential MYC binding sites in a wide range of human cells.

• Vector Backbone:

  lentiCRISPR v2 - Expresses Cas9

• Depositing Labs: Agnieszka Dzikiewicz-Krawczyk
• Publication: Kazimierska et al bioRxiv 2021.08.02.454734. doi: 10.1101/2021.08.02.454734.

Library Details

• Species: Human
• Genes targeted: 24,981 E-boxes
• gRNAs: 46,354
• Controls: 4 sgRNAs targeting MYC from the Brunello library (positive control)
  1,000 non-targeting sgRNAs from the Brunello library (negative control)
• 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: 100 ng/µL

Resource Information

• Protocols:
  • Library Amplification Protocol (119 KB)
• Depositor Data:
  • List of targeted E-boxes and gRNA sequences (3.9 MB)
  • Table of NGS Read Counts (1.2 MB)
  • NGS Read Distribution Histogram (180 KB)
• Terms and Licenses:
  • UBMTA

Depositor Comments

For enumeration of sgRNA read counts, we used the python script described in:
Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. PMID 28333914

Further analysis can be performed with various algorithms dedicated for CRISPR screens, such as MAGeCK, EgdeR, DeSeq2. We used DeSeq2 as it allows analysis of genes/sequences targeted by one or multiple sgRNAs.

NOTE: Lentiviral vectors can recombine during library amplification, resulting in a ~1.5kb band (may run differently in uncut samples) containing the origin and ampicillin resistance cassette. This recombination product should not adversely affect library function as it does not contain lentiviral packaging sequences or the PuroR gene and will not be selected during screening. If a strong, recombined band is observed after amplification, we recommend starting with a larger prep (gigaprep) from the original sample or gel purifying the amplified library to remove the recombinant band and performing another transformation with the purified sample.