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Fig. 3 | Genome Biology

Fig. 3

From: The collateral activity of RfxCas13d can induce lethality in a RfxCas13d knock-in mouse model

Fig. 3

RfxCas13d-mediated lethality may result from the collateral activity of RfxCas13d rather than off-target effects. a RT-qPCR to measure the knockdown efficiency of tdTomato crRNAs (n=3) in N2a cells stably expressing tdTomato (N2a-td). b Transcriptome analysis of N2a-td cells 48 h after transfection of plasmids encoding RfxCas13d and tdTomato crRNAs (n=3) or NT crRNA (n=3). c Body weight change curve of LSLRfxCas13d+/flAi14+/fl mice after injection of AAV-PHP.eB carrying tdTomato crRNAs (n=4) or NT crRNA (n=4). d Survival curve of LSLRfxCas13d+/flAi14+/fl mice after injection of AAV-PHP.eB carrying tdTomato crRNAs (n=4) or NT crRNA (n=4). e Western blot to measure tdTomato expression level in brain lysates of LSLRfxCas13d+/flAi14+/fl mice injected with AAV-PHP.eB carrying tdTomato crRNAs or NT crRNA at 12 dpi. f Schematic illustration of delivering AAV-PHP.eB carrying tdTomato crRNAs or NT crRNA into LSLRfxCas13d+/flAi14+/fl and LSLRfxCas13dfl/fl mice, and the possible outcomes. g Body weight change curve of LSLRfxCas13dfl/fl and LSLRfxCas13d+/flAi14+/fl mice after injection of AAV-PHP.eB carrying tdTomato crRNAs (n=4) or NT crRNA (n=4). h Survival curve of LSLRfxCas13dfl/fl and LSLRfxCas13d+/flAi14+/fl mice after injection of AAV-PHP.eB carrying tdTomato crRNAs (n=4) or NT crRNA (n=4). i Body weight change curve of Ai14 (n=4) and LSLRfxCas13d+/flAi14+/fl (n=3) mice after injection of AAV-PHP.eB carrying tdTomato crRNA 7. j Survival curve of Ai14 (n=4) and LSLRfxCas13d+/flAi14+/fl (n=3) mice after injection of AAV-PHP.eB carrying tdTomato crRNA 7. a One-way ANOVA with Dunn’s multiple comparisons test. g, j Two-tail unpaired t test. d, h, j Log-rank 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

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