Fig. 1
From: The collateral activity of RfxCas13d can induce lethality in a RfxCas13d knock-in mouse model
Mice died when knocking down Sik3-S in neurons using RfxCas13d. a Schematic illustration of RfxCas13d-mediated specifically knockdown of Sik3-S not Sik3-L mRNA. Sik3-L: a transcript encoding a long isoform of SIK3. b RT-qPCR to measure the knockdown efficiency of Sik3-S crRNAs (n=3). c Transcriptome analysis in N2a cells 48 h post-transfection of plasmids encoding RfxCas13d and Sik3-S crRNAs (n=3) or NT crRNA (n=3). d Workflow of knocking down Sik3-S in the adult mouse brain neurons. e Body weight change curve of LSLRfxCas13dfl/fl mice after injection of AAV-PHP.eB carrying Sik3-S crRNAs (n=5) or NT crRNA (n=5). f Survival curve of LSLRfxCas13dfl/fl mice after injection of AAV-PHP.eB carrying Sik3-S crRNAs (n=5) or NT crRNA (n=5). g, h Western blot and RT-qPCR to measure SIK-S/Sik3-S expression level in brain lysates of LSLRfxCas13dfl/fl mice injected with AAV-PHP.eB carrying Sik3-S crRNAs (n=3) or NT crRNA (n=3) at 20 dpi. b, h One-way ANOVA with Dunn’s multiple comparisons test. e Two-tail unpaired t test. f 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