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- W4249215296 abstract "Abstract High-throughput essentiality screens have enabled genome-wide assessments of the genetic requirements for growth and survival of a variety of bacteria in different experimental models. The reliance in many of these studies on transposon (Tn)-based gene inactivation has, however, limited the ability to probe essential gene function or design targeted screens. We interrogated the potential of targeted, large-scale, pooled CRISPR interference (CRISPRi)-based screens to extend conventional Tn approaches in mycobacteria through the capacity for positionally regulable gene repression. Here, we report the utility of the “CRISPRi-Seq” method for targeted, pooled essentiality screening, confirming strong overlap with Tn-Seq datasets. In addition, we exploit this high-throughput approach to provide insight into CRISPRi functionality. By interrogating polar effects and combining image-based phenotyping with CRISPRi-mediated depletion of selected essential genes, we demonstrate that CRISPRi-Seq can functionally validate Transcriptional Units within operons. Together, these observations suggest the utility of CRISPRi-Seq to provide insights into (myco)bacterial gene regulation and expression on a genome-wide scale." @default.
- W4249215296 created "2022-05-12" @default.
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- W4249215296 date "2018-06-29" @default.
- W4249215296 modified "2023-09-28" @default.
- W4249215296 title "CRISPRi-Seq for the Identification and Characterisation of Essential Mycobacterial Genes and Transcriptional Units" @default.
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- W4249215296 doi "https://doi.org/10.1101/358275" @default.
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