Supplementary Data for Genome-wide Characterization of Diverse Bacteriophages Enabled by RNA-Binding CRISPRi

Bacteriophages constitute one of the largest sources of unknown gene content in the biosphere. Even for well-studied model phages, robust experimental approaches to identify and study their essential genes remain elusive. We uncover and exploit the conserved vulnerability of the phage transcriptome to facilitate genome-wide protein expression knockdown via programmable RNA-binding protein dRfxCas13d (CRISPRi-ART) across diverse phages and their host. Establishing the first broad-spectrum phage functional genomics platform, we predict over 90 essential genes across four phage genomes, a third of which have no known function. These results highlight hidden infection strategies encoded in the most abundant biological entities on earth and provide a facile platform to study them.<br><b>Data S1. </b>Plasmids used in this study.<b>Data S2. </b>Bacterial strains used in this study.<b>Data S3. </b>Bacteriophages used in this study.<b>Data S4. </b>Oligonucleotides used in this study, not including oligo pools.<b>Data S5. </b>Data table describing metadata, raw and normalized read counts, crRNA fitness values.<b>Data S6. </b>Curated phage genome annotations for phages T4, T5, SUSP1, and PTXU04.<b>Data S7. </b>Plaque size measurements for all plaque assays in this study.<br>