Fig. 2
From: The collateral activity of RfxCas13d can induce lethality in a RfxCas13d knock-in mouse model
RfxCas13d-mediated lethality was not due to the loss of target gene function. a Western blot to measure SIK3-S expression level in brain lysates from Sik3-E5fl/fl and WT mice at 21 dpi. b Survival curve of Sik3-E5fl/fl (n=5) and WT (n=5) mice after injection of AAV-PHP.eB-hSYN-Cre. c Body weight change curve of Sik3-E5fl/fl (n=5) and WT (n=5) mice after injection of AAV-PHP.eB-hSYN-Cre. d, e RT-qPCR to measure the knockdown efficiency of Map2 (n=3) and Mapt (n=3) crRNAs in N2a cells. f Western blot to measure the knockdown efficiency of Rbfox3 crRNAs by knocking down overexpressed NeuN in HEK293T cells using RfxCas13d. g–i Transcriptome analysis in N2a cells 48 h after transfection of plasmids encoding RfxCas13d and crRNAs (n=3). j–l Body weight change curve of LSLRfxCas13dfl/fl mice after infection of PHP.eB carrying NT crRNA (n=5), Map2 crRNAs (n=5), Mapt crRNAs (n=5), or Rbfox3 crRNAs (n=5). m–o Survival curve of LSLRfxCas13dfl/fl mice after injection of AAV-PHP.eB carrying NT crRNA (n=5), Map2 crRNAs (n=5), Mapt crRNAs (n=5), or Rbfox3 crRNAs (n=5). p–r Western blot to measure MAP2 (at 6 dpi), Tau (at 25 dpi), or NeuN (at 50 dpi) expression level in brain lysates of LSLRfxCas13dfl/fl mice after injection of AAV-PHP.eB carrying NT crRNA, Map2 crRNAs, Mapt crRNAs or Rbfox3 crRNAs. d, e One-way ANOVA with Dunn’s multiple comparisons test. j–l Two-tail unpaired t test. b, m-o Log-rank test. Significance levels are noted as *P < 0.05, **P < 0.01, ****P < 0.0001 or ns (P > 0.05). All values are presented as mean ± SEM