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• W2804657653 abstract "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 ." @default.
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• W2804657653 date "2018-05-15" @default.
• W2804657653 modified "2023-09-30" @default.
• W2804657653 title "A CRISPRi screen in E. coli reveals sequence-specific toxicity of dCas9" @default.
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• W2804657653 doi "https://doi.org/10.1038/s41467-018-04209-5" @default.
• W2804657653 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5954155" @default.
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