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Purpose
This lentiviral CRISPR knockout library targets 182 core intrinsic immune evasion genes using sgRNAs selected from the mTKO library. It is designed for focused loss-of-function screening against a core set of genes shown to increase sensitivity or resistance of cancer cells to cytotoxic T lymphocyte (CTL) -mediated toxicity.
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Vector Backbone
pLCKO2 - does not express Cas9
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Depositing Labs
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Pooled Library | 159391 | gRNA pooled library† | 1 | $ 380 | Add to Cart | |
† A Cas9 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with Lenti-Cas9-2A-Blast (Addgene #73310) or any other plasmid or cell line stably expressing Cas9.
Library Details
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SpeciesMouse
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Genes targeted182
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gRNAs1,656
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Controls728 control gRNAs target 182 genes with no significant immune evasion fitness profile
100 gRNA target LacZ, EGFP, and luciferase
100 gRNA targeting intergenic regions -
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∼20 µL
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Concentration25 ng/µL
Resource Information
Depositor Comments
The depositing lab recommends using Bayestian Analysis of Gene Essentiality (BAGEL) to analyze fold changes.
MAGeCK/MAGeCK-VISPR algorithms can also be used for analysis.
The DrugZ algorithm is recommended for drug screens.
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.
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For your Materials & Methods section:
Mouse Validation (mVAL) library was a gift from Jason Moffat (Addgene #159391) -
For your References section:
Functional genomic landscape of cancer-intrinsic evasion of killing by T cells. Lawson KA, Sousa CM, Zhang X, Kim E, Akthar R, Caumanns JJ, Yao Y, Mikolajewicz N, Ross C, Brown KR, Zid AA, Fan ZP, Hui S, Krall JA, Simons DM, Slater CJ, De Jesus V, Tang L, Singh R, Goldford JE, Martin S, Huang Q, Francis EA, Habsid A, Climie R, Tieu D, Wei J, Li R, Tong AHY, Aregger M, Chan KS, Han H, Wang X, Mero P, Brumell JH, Finelli A, Ailles L, Bader G, Smolen GA, Kingsbury GA, Hart T, Kung C, Moffat J. Nature. 2020 Oct;586(7827):120-12. doi: 10.1038/s41586-020-2746-2. PubMed 32968282