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Bikard Lab - EcoWG1: E. coli Genome-wide Inhibition Library
(Pooled Library #131625)

  • Purpose

    This library contains ~20,000 unique guide RNAs targeting the non-template strand of every open reading frame and annotated small RNAs in the chromosome of E. coli MG1655. The library contains an average of 5 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). The psgRNA_EcoWG1 Library has the same backbone as the psgRNA Library, but the gRNAs in this library have have higher inhibition efficiency.

  • Vector Backbone

    psgRNA - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 131625 psgRNA_EcoWG1 1 $350 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 Strains LC-E18 (Addgene #115924) or LC-E75 (Addgene #115925).

Library Details

  • Species
    Escherichia coli MG1655
  • Genes targeted
    4,444
  • gRNAs
    21,417
  • Controls
    101

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:

    EcoWG1: E. coli Genome-wide Inhibition Library was a gift from David Bikard (Addgene #131625)
  • For your References section:

    On-target activity predictions enable improved CRISPR–dCas9 screens in bacteria. Calvo-Villamañán A, Ng JW, Planel R, Ménager, H, Chen A, Cui L, Bikard D. Nucleic Acids Res. 2020 Apr 30. pii: 5827197. doi: 10.1093/nar/gkaa294. PubMed 32352514