Fig. 8

RT-qPCR analysis of selected M1Cam and 81–176 CRISPRi transformants targeting flgR, flaA, flaB and flaA/flaB. RT-qPCR was performed on M1Cam flgR_913 (A), flaA_761 (B), flaB_1592 (C), flaAB_1034 (D), 81–176 flgR_559 (E) and flaAB_428 (F) to gain an insight into levels of repression elicited in each gene. RT-qPCR products lying upstream (U) and downstream (D) of the sgRNA target locations were designed for each gene, denoted as gene_U and gene_D respectively. The genes flaA and flaB were included in RT-qPCR analyses for M1Cam flgR_913 and 81–176 flgR_559 as downstream indicators of the consequences of flgR repression. * = p < 0.05, ** = p < 0.01 and **** = p < 0.0001. Statistics calculated using One-way T-tests of ∆∆ct values, N = 3 biological repeats, error bars = 95% CI