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Human Kinome CRISPR Knockout Pooled Libraries (Brunello)
(Pooled Libraries #1000000083, #75314, #75315, #1000000082, #75312, #75313)

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  • Purpose

    Human kinome CRISPR knockout pooled libraries. Each half-library (1–4 or 5–8) contains 3,052 unique sgRNAs targeting 763 human kinase genes for 4 guides per target.

  • Vector Backbone

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 1000000083 Two tubes, each with a half-library (gRNAs 1-4 or 5-8) in lentiCRISPR v2 
  • #75314 (gRNAs 1-4)
  • #75315 (gRNAs 5-8)
 1 $540 Add to Cart
Pooled Library 75314 One tube, contains gRNAs 1-4 in lentiCRISPR v2 1 $380 Add to Cart
Pooled Library 75315 One tube, contains gRNAs 5-8 in lentiCRISPR v2 1 $325 Add to Cart
Pooled Library 1000000082 Two tubes, each with a half-library (gRNAs 1-4 or 5-8) in lentiGuide‑Puro
  • #75312 (gRNAs 1-4)
  • #75313 (gRNAs 5-8)
 1 $540 Add to Cart
Pooled Library 75312 One tube, contains gRNAs 1-4 in lentiGuide-Puro 1 $380 Add to Cart
Pooled Library 75313 One tube, contains gRNAs 5-8 in lentiGuide-Puro 1 $325 Add to Cart
Individual Plasmids See below gRNAs 1-4 in lentiGuide-Puro 1 See below
Available to academic and nonprofits only.

Library Details

  • Species
    Human
  • Genes targeted
    763
  • gRNAs
    3,052
  • Controls
    100
  • 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.

Pooled libraries #75312, #75313, #75314, and #75315 will be delivered as a single tube containing a pooled half-library with 4 gRNAs per gene.

Pooled libraries #1000000082 and #1000000083 will be delivered as two tubes, each containing a pooled half-library with 4 gRNAs per gene (for a total of 8 unique gRNAs per gene).

  • Volume
    ∼10 µL
  • Concentration
    10 ng/µL

Resource Information

Addgene Notes

Screens can be performed using 4 individual guides per gene in 1-vector format (#75314 or #75315) or 2-vector format (#75312 or #75313). Individual 4 gRNA libraries can also be combined for larger screens (#1000000083 or #1000000082).

Note, the combined 8 gRNA library will provide 2x number of gRNAs per gene, but will also require a greater number of cells.

The amplification protocol provided, which recommends using 400 ng of DNA per transformation, was written in reference to the whole genome-wide libraries. These kinome-specific sub-pools contain fewer plasmids and thus require less DNA to achieve complete representation. Therefore, using the 100 ng of DNA as provided for transformation is sufficient.

NOTE: Lentiviral vectors can recombine during library amplification, resulting in an ~1 kb band containing the origin and ampicillin resistance cassette. This recombination product should not adversely affect library function as it does not contain lentiviral packaging sequences or the PuroR gene and will not be selected during screening. If this 1 kb band is observed after amplification, we recommend starting with a larger prep (gigaprep) from the original sample or gel purifying the amplified library to remove the recombinant band and performing another transformation with the purified sample.

Individual plasmids for human kinome CRISPR knockout library (guides 1–4) (Back to top)

Purpose

Individual gRNA plasmids. These gRNAs are represented in pooled library #75312 (guides 1–4) in lentiGuide-Puro. The depositors have also included 3 control gRNAs targeting EGFP.



Note! A separate Cas9 plasmid will need to be ordered to use these plasmids. Can be used in conjunction with lentiCas9-Blast (Addgene #52962) or otherwise with cell lines already expressing Cas9.

Description

  • Pricing & Shipping
    See individual plasmid page for shipping format and pricing.

gRNA plasmids targeting EGFP (controls)

ID Name Description
80034 EGFP gRNA (BRDN0000561167) 3rd generation lentiviral gRNA plasmid targeting EGFP
80035 EGFP gRNA (BRDN0000562805) 3rd generation lentiviral gRNA plasmid targeting EGFP
80036 EGFP gRNA (BRDN0000563266) 3rd generation lentiviral gRNA plasmid targeting EGFP

gRNA plasmids from human kinome CRISPR knockout library (guides 1–4)

To find a gene, enter the Entrez gene symbol in the search box. Click on an Addgene ID to access more information on each plasmid.

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:

    Human Kinome CRISPR pooled library (Brunello) was a gift from John Doench & David Root (Addgene #1000000083)
    Human Kinome CRISPR pooled library (Brunello) was a gift from John Doench & David Root (Addgene #75314 ; http://n2t.net/addgene:75314 ; RRID:Addgene_75314)
    Human Kinome CRISPR pooled library (Brunello) was a gift from John Doench & David Root (Addgene #75315 ; http://n2t.net/addgene:75315 ; RRID:Addgene_75315)
    Human Kinome CRISPR pooled library (Brunello) was a gift from John Doench & David Root (Addgene #1000000082)
    Human Kinome CRISPR pooled library (Brunello) was a gift from John Doench & David Root (Addgene #75312 ; http://n2t.net/addgene:75312 ; RRID:Addgene_75312)
    Human Kinome CRISPR pooled library (Brunello) was a gift from John Doench & David Root (Addgene #75313 ; http://n2t.net/addgene:75313 ; RRID:Addgene_75313)

  • For your References section:

    Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE. Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. PubMed 26780180
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