CRISPRi Library
(Pooled Library #62217)
- Purpose
Note: This library has been replaced by an updated version (see Depositor Comments below).
Human genome-scale CRISPRi library for inhibition of gene transcription. Expresses a gRNA from the U6 promoter and a puromycin resistance cassette and BFP from the EF1Alpha promoter.
- Vector Backbone
pSICO derivative - does not express Cas9
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
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Pooled Library | 62217 |
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1 |
† 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
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SpeciesHuman
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Genes targeted15,977
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gRNAs206,421
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Controls11,219
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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∼20 µL
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Concentration50 ng/µL
Resource Information
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Protocols
- Cell line creation and validation primer (Link opens in a new window)
- CRISPRi/a custom library cloning protocol (Link opens in a new window)
- CRISPRi/a screen data analysis pipeline on GitHub (Link opens in a new window)
- For more protocols, please see the Weismann lab's CRISPRi/a resource page (Link opens in a new window)
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Depositor Data
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Academic/Nonprofit Terms
Terms and Licenses
Depositor Comments
Note: This item has been discontinued. The Weissman lab has released an updated version of the library.
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.
Plasmid pHR-SFFV-dCas9-BFP-KRAB (#46911) was used for stable expression of the dCas9-KRAB fusion protein for CRISPRi experiments in the associated publication. Plasmid pHR-SFFV-KRAB-dCas9-P2A-mCherry (#60954) is a non-inducible version of the KRAB-dCas9 construct described in Figure 5A of the paper.
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:
CRISPRi library was a gift from Jonathan Weissman (Addgene #62217)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. 10.1016/j.cell.2014.09.029 PubMed 25307932