Genome-wide Mouse Lentiviral CRISPR gRNA Library v1
(Pooled Library #50947)
- 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Note: This library has been replaced by an updated version (see below).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
- Depositing Labs
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
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Pooled Library | 50947 |
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1 |
† 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
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SpeciesMouse
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Genes targeted19,150
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gRNAs87,897
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Lentiviral Generation3rd
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.
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Volume∼50 µL
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Concentration20 ng/µL
Resource Information
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Protocols
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Depositor Data
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Terms and Licenses
Academic/Nonprofit Terms
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 26-bp 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 Cas9-expressing plasmid used in Koike-Yusa et al. 2014, pPB-LR5.1-EF1a-puro2ACas9, may be obtained from the Wellcome Trust Sanger Institute Archives (Link opens in a new window).
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. doi: 10.1038/nbt.2800. PubMed 24535568 (Link opens in a new window)