Teichmann Retroviral Mouse Genome-wide CRISPR Knockout Library
(Pooled Library #104861)

• Purpose:

  This retroviral CRISPR library targets 18,424 mouse genes using the gRNAs designed by the Kosuke Yusa Lab. The use of retrovirus enables this library to be used for cell lines that are poorly infected by lentivirus.

• Vector Backbone:

  pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP (Plasmid #102796) - does not express Cas9

• Depositing Labs: Sarah Teichmann
• Publication: Henriksson et al. Cell. 2019 Feb 5;176(4):882-896. 10.1016/j.cell.2018.11.044.

Library Details

• Species: Mouse
• Genes targeted: 18,424
• gRNAs: 90,230
• Virus Type: Retrovirus

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: ∼50 µL
• Concentration: 20 ng/µL

Resource Information

• Protocols:
  • Library Replication Protocol - Yusa Lab (DOCX, 23 KB)
• Depositor Data:
  • List of gRNA Sequences (XLSX, 4.2 MB)
  • Distribution of NGS Reads (CSV, 8.2 MB)
• Articles Citing this Pooled Library:
  • Retroviral Mouse Genome-wide CRISPR KO (#104861)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA
• Evrogen Limited Use Label License for FPs

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

The depositing lab recommends 3–5 electroporations for amplification.

NOTE: Retroviral vectors can recombine during library amplification, resulting in an ~2 kb band containing the origin and ampicillin resistance cassette. This recombination product should not adversely affect library function as it does not contain retroviral packaging sequences or mammalian selection markers. If this 2 kb band forms the predominant product 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.