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. 2019 Sep 24;12(5):511–528. doi: 10.1007/s12195-019-00598-9

Figure 7.

Figure 7

Duty cycle increases repression and expression variability in strains carrying the dCas9-mCherry-LINuS repressor. (a) Strains carrying dCas9-mCherry-LINuS and the sgTEF1 guide RNA were exposed to cycles of 15 µW/cm2 or 135 µW/cm2 blue light at increasing duty cycle (0% off 5% 1 s on/19 s off, 25% 5 s on/15 s off, 50% 10 s on 10 s off 75% 15 s on 5 s off 100% on). Increasing duty cycle increases repression up to a maximum of 1.5-fold. Error bars represent 95% confidence intervals on the average of the median fluorescence from n = 3 biological replicates. (b) Duty cycle increases the population of low expressing cells while leaving a significant fraction of the population distributed through the wild-type (dark) expression levels. Dashed lines represent the first and third quartile for cells carrying dCas9-mCherry-LINuS/sgTEF1 in the dark. (c) Plotting the subpopulation ratio (NL/NH) shows that increasing light dosage increases the ratio of low expressors to high expressors in the population (up to 6-fold for dCas9-mCherry-LINuS-Mxi1).