Human Mitochondrial Knockout (mtKO) gRNA Library
(Pooled Library #247670)

• Purpose:

  The mtKO guide RNA library targets key mitochondria-associated genes across major pathways. It enables pooled screens to identify mitochondrial dependencies in normal and malignant cells.

• Vector Backbone:

  pUSEPR (GB, 16 KB) - does not express Cas9

• Depositing Labs: Chunzhang Yang
• Publication: Kaur et al. Proc Natl Acad Sci U S A. 2025 Jul 22;122(29):e2502285122. doi: 10.1073/pnas.2502285122.

Library Details

• Species: Human
• Genes targeted: 1,034
• gRNAs: 17,523
• Controls: 1,409 core essential genes (8,454 guides). 405 non-essential genes (2,430 guides). 435 guides for non-targeting intergenic regions.
• 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:
  • Electroporation and Recovery Protocol (DOCX, 19 KB)
  • PCR primer details can be found in the supplementary information appendix in the publication. Expected amplicon sizes:
    • PCR1: ~600 bp
    • PCR2: 200–300 bp
• Depositor Data:
  • List of guides (XLSX, 560 KB)
  • Guide Distribution (PNG, 326 KB)
• Scripts:
  • MAGeCK , BAGEL2 , Drugz . The code and script should be customized depending on the experimental design. The depositing lab recommends reaching out to them directly for specific analysis.

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.