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Addgene

Genome-wide Mouse Lentiviral CRISPR gRNA Library v1
(Pooled Library #50947)

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  • Purpose

    Note: This library has been replaced by an updated version (see below).

    A gRNA library against 19,150 mouse protein coding genes, designed for use in lentivirus.

  • Vector Backbone

    pKLV-U6gRNA(BbsI)-PGKpuro2ABFP- does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 50947

  • gRNA pooled library†

1
Available to Academic and Nonprofits Only

  • † A Cas9 plasmid is NOT included with this item and will have to be ordered separately, or be used with cell lines already expressing Cas9.

Library Details

  • Species
    Mouse
  • Genes targeted
    19,150
  • gRNAs
    87,897
  • Lentiviral Generation
    3rd

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
    ∼50µL
  • Concentration
    20ng/µL

Resource Information

Depositor Comments

Note: This item has been discontinued. The Yusa lab has released an updated version of the library here.

gRNA library construction. A pool of 87,897 79-mer oligonucleotides was purchased from CustomArray Inc. They were converted into double-stranded DNA by PCR and 26bp fragments were excised by BbsI digestion and purified from a PAGE gel. Transformation of DH10B E. coli with the ligated DNA yielded a plasmid library derived from approximately 15 million E. coli colonies. The gRNA is immediately downstream of the U6 promoter.

The construct containing Cas9, pPB-LR5.1-EF1a-puro2ACas9, may be obtained from the Wellcome Trust Sanger Institute Archives.

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 pooled libraries were described, and include Addgene in the Materials and Methods of your future publications.

  • Example for your Materials & Methods section:

    Genome-wide mouse lentiviral CRISPR gRNA library was a gift from Kosuke Yusa (Addgene #50947)

  • For your References section:

    Genome-wide recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library . Koike-Yusa H, Li Y, Tan EP, Velasco-Herrera Mdel C, Yusa K. Nat Biotechnol. 2014 Mar;32(3):267-73. doi: 10.1038/nbt.2800. Epub 2013 Dec 23. 10.1038/nbt.2800PubMed 24535568