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

• Depositing Labs: David Bikard
• Publication: Calvo-Villamanan et al Nucleic Acids Res. 2020 Apr 30. pii: 5827197. doi: 10.1093/nar/gkaa294. -->

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

Resource Information

• Protocols:
  • Library Transformation Protocol (17 KB)
  • Illumina Screening Protocol (15 KB)
  • Illumina Sequencing Protocol (20 KB)
• Depositor Data:
  • List of gRNAs (2.7 MB)
  • NGS Read Counts (743 KB)
• Terms and Licenses:
  • UBMTA

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.