 |
Journal articles
A CRISPRi screen in E. coli reveals sequence-specific toxicity of dCas9
Abstract : High-throughput CRISPR-Cas9 screens have recently emerged as powerful tools to decipher gene functions and genetic interactions. Here we use a genome-wide library of guide RNAs to direct the catalytically dead Cas9 (dCas9) to block gene transcription in Escherichia coli. Using a machine-learning approach, we reveal that guide RNAs sharing specific 5-nucleotide seed sequences can produce strong fitness defects or even kill E. coli regardless of the other 15 nucleotides of guide sequence. This effect occurs at high dCas9 concentrations and can be alleviated by tuning the expression of dCas9 while maintaining strong on-target repression. Our results also highlight the fact that off-targets with as little as nine nucleotides of homology to the guide RNA can strongly block gene expression. Altogether this study provides important design rules to safely use dCas9 in E. coli.
Cited literature [35 references]
https://hal-pasteur.archives-ouvertes.fr/pasteur-01819630
Contributor : Corinne Billion <>
Submitted on : Wednesday, June 20, 2018 - 4:53:57 PM
Last modification on : Monday, December 14, 2020 - 3:35:20 PM
Long-term archiving on: : Monday, September 24, 2018 - 11:42:46 PM
Contributor : Corinne Billion <>
Submitted on : Wednesday, June 20, 2018 - 4:53:57 PM
Last modification on : Monday, December 14, 2020 - 3:35:20 PM
Long-term archiving on: : Monday, September 24, 2018 - 11:42:46 PM
File
s41467-018-04209-5.pdf
Publication funded by an institution
Licence
Distributed under a Creative Commons Attribution 4.0 International License