Fig. 5: Use of a multi-guide array. | Communications Biology

Fig. 5: Use of a multi-guide array.

From: Cas13b-dependent and Cas13b-independent RNA knockdown of viral sequences in mosquito cells following guide RNA expression

Fig. 5

a Pol III promoter (U6)-driven RNA guide arrays used to test the effect of guide position on knockdown. Constructs P1, P2 and P3 have the active guide (guide 2) in positions 1, 2 and 3, respectively, of the three-position array, with guides against an absent target (AmCyan) in the remaining positions. A mock array (PC) with all the positions filled with non-targeting guides (AmC1, AmC2, AmC3) was used as control. b–e Interference of the CHIKV-luciferase reporter with U6 promoter-driven RNA array in Aag2 (b), AF05 (c), Dcr2 knockout AF319 (d) and C6/36 (e) cells. Each bar represents the mean values of firefly luciferase activity normalised to the Renilla luciferase activity (FF/RL); the whiskers are the standard deviation from three independent transfections with four to six replicates each and each dot represents a replicate. b–d Linear mixed effects model, e two-way ANOVA. ***P < 0.001, **P < 0.01, *P < 0.05, ns: not significant (Supplementary Tables 16–19). DR: non-variable RNA guide backbone, U6T: U6 promoter terminator, P1: position 1, P2: position 2, P3: position 3.

Back to article page