Figure 6.

A plasmid transcript targeted by Cas13a and expressed under the threshold can be tolerated and can confer benefits to the host. See also Fig. S7.

(A) Experimental setup for evaluating tolerance for a plasmid-expressed beneficial gene. The two targets (K1, K2) fall within the kan^R transcript, which is expressed under different constitutive promoters (P2, P5, P8).

(B) Impact of Cas13a-based targeting of the kan^R transcript expressed under the different constitutive promoters. Bars represent the mean of quadruplicate independent experiments (n = 4). Statistical significance was calculated by comparing the transformation fold-reduction to that without the kan^R gene. ***: p < 0.001. **: p < 0.01. *: p < 0.05. ns: not significant.

(C) Heat map comparing growth in the presence of kanamycin (10 μg/mL) or hygromycin (100 μg/mL) for constructs associated with a low transformation fold-reduction in B. Values represent the average turbidity after 12 hours of growth from 9 biological replicates. Constructs lacking the kan^R gene serve as negative controls, while the non-targeting crRNA plasmid serves as a non-targeting (NT) control. See Fig. S7 for the same growth measurements with a higher concentration of kanamycin (50 μg/mL).