Extended Data Fig. 1: Optimization of ASO treatment.
From: Programmable antisense oligomers for phage functional genomics
a. Replication cycle of ΦKZ. b. Scheme depicting the optimization for the CFU/PFU assay (top) and assessment of ASO toxicity (bottom). c. PAO1 cells were pretreated with 6 µM ASO against chmA linked to the carrier peptides (RXR)4XB, (KFF)3K or TAT for 30 min. Cells were infected with ΦKZ at an MOI = 0.0001 and incubated for 180 min followed by CFU/PFU determination. Representative result of two independent experiments. d. PAO1 cells were pretreated with 6 µM ASO with indicated lengths and target sequences against RBS and AUG of chmA for 30 min. Cells were infected with ΦKZ at an MOI = 0.0001 and incubated for 180 min followed by CFU/PFU determination. SD, Shine-Dalgarno sequence. Representative result of three independent experiments. e. PAO1 cells were pretreated with 6 µM ASO against chmA for 30, 15, 10 or 5 min. Cells were infected with ΦKZ at an MOI = 0.0001 and incubated for 180 min followed by CFU/PFU determination. f. PAO1 cells were pretreated with 6 µM ASO against chmA for 30 min. Cells were infected with ΦKZ at an MOI = 0.0001 and incubated for 30, 70, 140, and 210 min followed by CFU/PFU determination. g. PAO1 cells were pretreated with 3 to 18 µM ASO against chmA for 30 min. Cells were incubated for 180 min followed by CFU/PFU determination. h. PAO1 cells were pretreated with 0.25 to 6 µM ASO against chmA for 30 min. Cells were infected with ΦKZ at an MOI = 5 and incubated for 180 min followed by CFU/PFU determination. i. PAO1 cells were pretreated with 6 µM ASO against chmA for 30 min. Cells were infected with ΦKZ at an MOI between 0.001 and 1 and incubated for 30, 70, 210 min followed by CFU/PFU determination.
