Human Genome-Wide Dual sgRNA CRISPRi v3 (LGR Backbone)
(Pooled Library #254658)

• Purpose:

  The library may be used for pooled, genome-wide CRISPR screens in human cells expressing the correct CRISPR machinery. The library contains gRNAs for inhibition (CRISPRi) of protein coding genes in the human genome for packaging and delivery by lentivirus. This library uses the dual sgRNA design reported by Replogle et al. (2022) for hCRISPRi_dual (Addgene #187246), but is delivered in a different backbone.

• Vector Backbone:

  pLGR_dual_guide-Puro-mycNLS-BFP backbone (PDF, 217 KB; PNG, 327 KB) - does not express dCas9

• Depositing Labs: Alexander Brown, Laboratory for Genomics Research (LGR)
• Publication: Laboratory for Genomics Research (LGR) - Human CRISPR Pooled Libraries

Library Details

• Species: Human
• Elements: 20,528 total dual-guide plasmids
• Genes targeted: 18,905
• Controls: 1,026 NTC dual sgRNA control elements
• 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.

• Volume: ∼20 µL
• Concentration: 50 ng/µL

Resource Information

• Protocols:
  • Library Amplification Protocol (PDF, 463 KB)
  • PCR Protocol (DOCX, 20 KB)
• Depositor Data:
  • List of gRNA sequences (TSV, 4.2 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

Growth issues were noticed when using pre-mixed media and LB plates. Because of this, we recommend using MegaX DH10B™ T1R Electrocomp™ Cells (Invitrogen #C6400-03) cultured at 37°C, which is what we use to generate all our libraries. Additionally, we recommend switching from ampicillin to carbenicillin to prevent satellite colonies.