Mouse Genome-wide CRISPRi v2 Library
(Pooled Libraries #1000000092, #83987)

• Purpose:

  Version 2 - Mouse genome-scale CRISPRi library for inhibition of gene transcription. Available as 5 gRNAs or 10 gRNAs per gene.

• Vector Backbone:

  pCRISPRia-v2 (Plasmid #84832) - does not express Cas9

• Depositing Labs: Jonathan Weissman
• Publication: Horlbeck et al Elife. 2016 Sep 23;5. pii: e19760. doi: 10.7554/eLife.19760

Library Details

• Species: Mouse
• Genes Targeted: 20,003
• gRNAs:
  
  • #83987: 107,415
  • #1000000092: 214,830
• Controls:
  
  • #83987: 2,170
  • #1000000092: 4,340
• Lentiviral Generation: 3rd

Library Shipping

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

Pooled Library #83987 will be delivered as a single tube containing a pooled half-library with 5 gRNAs per gene.

Pooled Library #1000000092 will be delivered as two tubes, each containing a pooled half-library with 5 gRNAs per gene (for a total of 10 unique gRNAs per gene).

• Volume: ∼20 µL
• Concentration: 25 ng/µL

Resource Information

• Protocols:
  • Illumina Sequencing Sample Preparation (PDF, 752 KB)
  • Liquid Culture Re-Amplification Protocol (PDF, 70 KB)
  • Cell Line Creation and Validation Primer (PDF, 125 KB)
  • CRISPRi/a Custom Library Cloning Protocol (PDF, 92 KB)
  • For more links and protocols, please see the Weissman lab's CRISPRi/a screening materials page
• Depositor Data:
  • Guide Sequences (ZIP, 16 MB)

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

In addition to dCas9-KRAB (#46911), another dCas9-containing construct that can be used with these libraries is: pHR-SFFV-KRAB-dCas9-P2A-mCherry (#60954).

The data analysis pipeline for analyzing sequencing data is available on GitHub.