Fig. 6: Programmable transcriptome-scale translational suppression unveils the gene fitness landscape of diverse phages.
From: CRISPRi-ART enables functional genomics of diverse bacteriophages using RNA-binding dCas13d
a, Transcriptome-wide fitness landscape of phages T5 (162 coding sequences), SUSP1 (138 coding sequences) and PTXU04 (92 coding sequences). The outer track displays the phage gene map with genes categorized by function, and the inner track shows fitness measured for each corresponding gene, coloured by Fit (red), Semi-fit (orange) or Not fit (blue). Gene fitness is shown as the mean of 3 biological replicates. b, Distribution of phage gene annotation quality for all phages across the full genome (top) and Fit genes (bottom). c, Phage-gene fitness for putative polyvalent protein encoded by PTXU04_00025 and csrA homologue PTXU04_00028. crRNA fitness is shown as the median of 3 biological replicates. d, Corresponding plaque assays validating pooled gene fitness classification calls with single crRNAs. Targeting Fit gene PTXU04_00025 (gp25) causes a large EOP reduction, while targeting Semi-fit gene PTXU04_00028 (gp28) leads to a reduction in plaque size. -con is a non-targeting crRNA control. Representative plaque assays of 3 independent replicates are shown.
