@article{DeWet2018,
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 {\&}{\#}039;CRISPRi-Seq{\&}{\#}039; 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.},
author = {de Wet, Timothy J and Gobe, Irene and Mhlanga, Musa M and Warner, Digby F},
doi = {10.1101/358275},
journal = {bioRxiv},
keywords = {OA,fund{\_}not{\_}ack,original},
mendeley-tags = {OA,fund{\_}not{\_}ack,original},
month = {jan},
pages = {358275},
title = {{CRISPRi-Seq for the identification and characterisation of essential mycobacterial genes and transcriptional units}},
url = {http://biorxiv.org/content/early/2018/06/29/358275.abstract},
year = {2018}
}

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