Bikard E. coli Genome-wide Inhibition Library
(Pooled Library #115927)
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.
psgRNA - does not express Cas9
|Item||Catalog #||Description||Quantity||Price (USD)|
|Pooled Library||115927||psgRNA Library†||1||$500||Add to Cart|
SpeciesEscherichia coli MG1655
Controls1 - backbone is used as a control
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.
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.
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
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