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. 2020 Feb 21;64(3):e01789-19. doi: 10.1128/AAC.01789-19

FIG 2.

FIG 2

Performance of the CRISPRi system in alleviating the class 1 integron-mediated antibiotic resistance in E. coli. (A) Schematic diagram of a CRISPRi system to attenuate the antibiotic resistance caused by the class 1 integron. (B and E) Transcription of the dfrB2 cassette (B) or sul1 (E) in recombinant E. coli strains in the presence or absence of aTc. (C and F) Determination of the IC50 of TMP (C) or SUL (F) in recombinant E. coli with and without aTc induction. (D and G) Inhibition of fluorescence in MABA at different concentrations of TMP (D) or SUL (G) with aTc induction. (H) Correlation analysis between the transcription of the dfrB2 cassette and the IC50 of TMP. (I) Correlation analysis between the sul1 transcription and the IC50 of SUL. (J) Correlation analysis of the transcriptional levels of the dfrB2 cassette and sul1. aTc, anhydrotetracycline; IC50, half-maximal inhibitory concentration; TMP, trimethoprim; SUL, sulfamethoxazole; MABA, microplate alamarBlue assay; R(0–6), E. coli C600[R388/plv-dCas9-R(0–6)]; R388, E. coli C600(R388); ATCC, E. coli ATCC 25922. All data represent the means ± standard deviations of biological triplicates. *, P < 0.05; **, P < 0.01; ***, P < 0.001.