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Bovine CRISPR Knockout Libraries
(Pooled Library #213927, #213928)

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  • Purpose

    These are two genome-wide CRISPR knockout libraries targeting the cattle protein-coding genome. One uses lentivirus to deliver into cells (LV2btCRISPRko.v1), and the other uses the PiggyBac transposon system to deliver the library into hard-to-transduce cells (PBbtCRISPRko.v1). These are identical CRISPR libraries with the same inserts but different vector backbones for two methods of library delivery.

  • Vector Backbone

    pKLV2-U6gRNA5(BbsI)-PGKpuro2ABFP-W - does not express Cas9

    PB_U6gRNA5(BbsI)-PGKPuro2ABFP-w (GB, 18 KB) - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 213927 LV2btCRISPRko.v1† 1 $640 Add to Cart
Pooled Library 213928 PBbtCRISPRko.v1† 1 $640 Add to Cart
Available to Academic and Nonprofits Only

  • † A Cas9 plasmid is NOT included with this item and will have to be ordered separately. The lentiCas9-Blast (#52962) or something similar that expresses wildtype spCas9 (codon optimized for mammalian cells) can be used to make cell lines express Cas9.

Library Details

  • Species
    Cow
  • Protein coding genes targeted
    21,165 (4.4 guides per gene on average)
  • Breakdown

    • Number of genes with 1 guide: 38
    • Number of genes with 2 guides: 71
    • Number of genes with 3 guides: 112
    • Number of genes with 4 guides: 11238
    • Number of genes with 5 guides: 9704
    • Number of genes with 6 guides: 2
    • Total: 94,000 targeting the coding protein genes
  • Controls
    2,000 non-cutting sgRNAs designed for the cow genome
  • Lentiviral generation
    2nd

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
    ∼20 µL
  • Concentration
    50 ng/µL

Resource Information

Depositor Comments

PBbtCRISPRko.v1, the PiggyBac version of the library, has not been sequenced. It was generated using the same oligo pool as LV2btCRISPRko.v1, and sufficient coverage was ensured during cloning. The quality of PBbtCRISPRko.v1 should be comparable to LV2btCRISPRko.v1.

How to cite this pooled library ( Back to top )

These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    Bovine CRISPR Knockout LV2btCRISPRko.v1 Library was a gift from Wenfang Spring Tan (Addgene #213927)
    Bovine CRISPR Knockout PBbtCRISPRko.v1 Library was a gift from Wenfang Spring Tan (Addgene #213928)

  • For your References section:

    GARP and EARP are required for efficient BoHV-1 replication as identified by a genome wide CRISPR knockout screen. Tan WS, Rong E, Dry I, Lillico SG, Law A, Digard P, Whitelaw B, Dalziel RG. PLoS Pathog. 2023 Dec 6;19(12):e1011822. doi: 10.1371/journal.ppat.1011822. PubMed 38055775
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