Human Genome-Wide CRISPRa v2 (LGR Backbone)
(Pooled Library #254657)

• 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 activation (CRISPRa) of protein coding genes in the human genome for packaging and delivery by lentivirus. This library uses the sgRNA design reported by Horlbeck et al. (2016) for hCRISPRa-v2 (Addgene #83978), but is delivered in a different backbone.

• Vector Backbone:

  pLGR002 LGR Cas9 guide vector (Plasmid #188320) - 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
• Genes targeted: 18,915
• gRNAs: 104,540
• Controls: 1,895 NTCs
• 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: ∼25 µL
• Concentration: 100 ng/µL

Resource Information

• Protocols:
  • Library Amplification Protocol (PDF, 463 KB)
  • PCR Protocol (DOCX, 19 KB)
• Depositor Data:
  • List of gRNA sequences (CSV, 5.9 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.