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Bikard E. coli Genome-wide Inhibition Library
(Pooled Library #115927)

  • Purpose

    This library contains ~93,000 unique guide RNAs targeting random positions along the genome of E. coli MG1655 with a NGG PAM motif. The library contains an average of 19 targets per gene and was made by cloning a pool of guide RNAs oligos into psgRNA (#114005) (ori: ColE1) by Gibson assembly.

    This library can be used with SpdCas9 to perform pooled CRISPRi screens as shown ( Cui et al., 2018 ; Rousset et al. 2018). Screens can be performed using strain LC-E75 (#115925), E. coli strain MG1655 expressing dCas9 under the control of a Tet promoter integrated at the 186 attB site, or strain FR-E01 (#118727), E. coli strain MG1655 expressing dCas9 under the control of a Tet promoter integrated at the HK022 attB site.

  • Vector Backbone

    psgRNA - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 115927 psgRNA Library 1 $500 Add to Cart
Available to Academic and Nonprofits Only
  • A dCas9-expressing strain is NOT included with this item and will have to be ordered separately. Can be used in Strain LC-E75 (Addgene #115925) or Strain FR-E01 (Addgene #118727).

Library Details

  • Species
    Escherichia coli MG1655
  • Genes targeted
    ~4,500
  • gRNAs
    92,919
  • Controls
    1 - backbone is used as a control

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.

NOTE: 100 ng of the library can be electroporated into cells for an experiment. Sufficient sample is provided for several experiments and amplification of the library.

  • Volume
    ∼20µL
  • Concentration
    50ng/µL

Depositor Comments

The depositing lab recommends using their code for analyzing Illumina sequencing results.

For additional information on use of this library, please see Rousset et al., 2018.

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:

    E. coli Genome-wide Inhibition library was a gift from David Bikard (Addgene #115927)
  • For your References section:

    A CRISPRi screen in E. coli reveals sequence-specific toxicity of dCas9. Cui L, Vigouroux A, Rousset F, Varet H, Khanna V, Bikard D. Nat Commun. 2018 May 15;9(1):1912. doi: 10.1038/s41467-018-04209-5. PubMed 29765036
    and
    Genome-wide CRISPR-dCas9 screens in E. coli identify essential genes and phage host factors. Rousset F, Cui L, Siouve E, Becavin C, Depardieu F, Bikard D. PLoS Genet. 2018 Nov 7;14(11):e1007749. doi: 10.1371/journal.pgen.1007749. eCollection 2018 Nov. PubMed 30403660