Fig. 5
From: Switching the activity of Cas12a using guide RNA strand displacement circuits
Transcriptional regulation via SD gRNAs and dCas12a in E. coli. a Repression of mVenus fluorescence by different versions of target-based SD gRNAs in E. coli with and without the expression of a cognate trigger. The design of the different versions is shown on the left. Version A is identical to the target-based SD gRNAs shown in Fig. 3. Expression of the dsDNase inactive mutant dCas12a is induced with IPTG and the fluorescence per OD at 4 h after induction is compared with the fluorescence per OD of uninduced cells containing the same SD gRNA plasmid and no cognate trigger. A regular dCas12a gRNA is used as a positive control for repression (ctrl: control trigger, t1: cognate trigger, reg: regular gRNA) (N = 4, t-distribution two-sided 90% confidence interval). b Fold repression by the SD gRNA version C design for four different target sequences on the mVenus gene at 4 h after induction (ctrl: control trigger, t1-t6: cognate triggers, reg: regular gRNA) (N = 4, t-distribution two-sided 90% confidence interval). c Crosstalk between different SD gRNAs and their triggers at 4 h after induction (N = 4). d Crosstalk between different SD gRNAs and their triggers at 4 h after induction normalized by the fold repression of the respective positive control gRNA. (N = 4) Source data are provided as a Source Data file
