Extended Data Fig. 8: Delivery efficiencies into pathogenic strains of E. coli and K. pneumoniae.
From: In situ targeted base editing of bacteria in the mouse gut
a) Delivery efficiency assays in MG1655 carrying TN03-OmpC of cosmids harboring A8 (orange line) or 1A2 (blue line) gpJ chimeras and P2 STF. Grey line shows a comparison of the delivery efficiency of gpJ chimera 1A2 into MG1655 carrying OmpC-EDL933. Transduction of a payload encoding a sfGFP fluorescent protein gene was measured in a flow cytometer (excitation: 488 nm, emission: 530/30 BP) at different MOIs. b) Flow cytometry histogram of E. coli TN03 after delivery with a cosmid harboring λ-A8 gpJ chimera and λ-WT STF (blue line) or λ-K5 STF (red line) at an estimated MOI of ~10. Transduction of a payload encoding a mCherry fluorescent protein gene was measured in a flow cytometer (excitation: 561 nm, emission: 620/15 BP). c) Delivery efficiency into E. coli TN03 of λ particles carrying the λ-K5 STF chimera and payload from panel b at different MOIs. d) Delivery efficiencies of a DNA payload encoding the fluorescent gene venus (plasmid p2075) into UTI89 (gpJ 1A2, λ-K1F STF chimera), or a DNA payload encoding the fluorescent gene mCherry (plasmid p2074) into TN03 (gpJ A8, λ-K5 STF chimera) or Klebsiella pneumoniae (gpJ A8, λ-KL106 STF chimera), measured by flow cytometry (excitation: 488 nm, emission: 530/30 BP).
