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. 2021 Feb 28;49(6):3584–3598. doi: 10.1093/nar/gkab126

Figure 3.

Figure 3.

TAP system specifically kills targeted recipients in a mix of targeted and non-targeted E. coli recipient cells. (A) Viable transconjugant cells and percentage of transconjugants (ratio T/R + T) through TAPkn-Cas9-nsp or TAPkn-Cas9-lacZ2 transfer from donor to a mixed lac+ and lac– recipient population. Two-tails unpaired t-test was performed on log10-transformed values. ns = non-significant P-value >0.05; ***P-value <0.0002. (B) Quantification of fold-increase in lac+ and lac– recipient populations counts over the 6h of mating. Mean and SD are calculated from 4 independent experiments. Donors: TAP-Cas9-nsp (LY1369) or TAP-Cas9-lacZ2 (LY1370); recipients lac+ (LY827) and lac– (LY848). Two-tails unpaired t-test was performed. ns = non-significant P-value >0.05; **P-value <0.0021. (C) Single-cell quantification showing cell length increase in the targeted lac+ transconjugant cells but not non-targeted lac- transconjugants. The time of TAP acquisition (red dashed line at 0 min) corresponds to a 15% increase in the green fluorescence in the transconjugant cells. Cell length average is indicated with SD (n cells analysed). Donor TAP-Cas9-lacZ2 (LY1380); recipients HU-mCherry lac+ (LY248) and DnaN-mCherry lac– (LY1423). Multiple t-test were performed corrected with Holm–Sidak method. Stars indicate the time with significant difference (P-value < 0.05). Significant difference was observed from this point until the end of the analysis.