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CRISPRa Library
(Pooled Library #60956)

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  • Purpose

    Note: This library has been replaced by an updated version (see Depositor Comments below).

    Human genome-scale CRISPRa library for activation of gene transcription. Expresses a gRNA from the U6 promoter and a puromycin resistance cassette and BFP from the EF1Alpha promoter.

  • Vector Backbone
    pU6-sgRNA EF1Alpha-puro-T2A-BFP (Plasmid #60955) - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 60956 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 otherwise used with cell lines already expressing Cas9.

Library Details

  • Species
    Human
  • Genes targeted
    15,977
  • gRNAs
    198,810
  • Controls
    5,968
  • Lentiviral Generation
    3rd

Library Shipping

Each library is delivered in microcentrifuge tubes on blue ice. A 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

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

The library should be amplified before attempting to create virus. The use of high efficiency, electrocompetent cells is strongly advised (e.g. NEB DH5a cells, Invitrogen DH5a cells, or Lucigen Endura). Follow manufacturer's suggested protocol for transformation.

To ensure the maintenance of library diversity, calculate your transformation efficiency. 30x coverage of the library is recommended.

Plasmids pHRdSV40-dCas9-10xGCN4_v4-P2A-BFP (#60903) and pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS (#60904) comprised the dCas9-SunTag and scFV-VP64 components used with the CRISPRa library, as described in the associated publication.

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:

    CRISPRa library was a gift from Jonathan Weissman (Addgene #60956)

  • For your References section:

    Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M, Weissman JS. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct 9. PubMed 25307932
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