Fig. 4

The collateral activity of RfxCas13d was positively correlated with the abundance of target RNA in mammalian cells. a Western blot to measure the expression level of RfxCas13d and tdTomato 24 h after transfection plasmids encoding RfxCas13d, tdTomato, and crRNAs into HEK293T cells. 3F-tdTomato means tdTomato with 3× Flag tag at N-terminal. b RT-qPCR to measure the RNA level of RfxCas13d in a. td represents tdTomato. EV represents empty vector. c, d RT-qPCR to measure the mRNA level of tdTomato (c) and RfxCas13d (d) 24 h after transfection of plasmids encoding RfxCas13d and crRNA into the inducible-expressing tdTomato HEK293T cells. The cells were pretreated with different concentrations of doxycycline for 16 h. e–g RT-qPCR to measure the knockdown efficiency of ACTB, LDHB, and YEHAE crRNAs in HEK293T cells (n=3). h–m RT-qPCR to measure the mRNA levels of RfxCas13d (h–j) and mCherry (k–m) in the HEK293T cells 24 h after transfection of plasmids respectively encoding RfxCas13d, mCherry, and crRNA. b–m One-way ANOVA with Dunn’s multiple comparisons test. Significance levels are noted as *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, or ns (P > 0.05). All values are presented as mean ± SEM