Fig. 5
From: Cas13b-mediated RNA targeted therapy alleviates genetic dilated cardiomyopathy in mice
Restoration of cardiac function in DCM through PspCas13b-mediated hTNNT2R141W knockdown. (A) HW/BW ratios of the WT (n = 13), R141W (n = 9) and R141W + PspCas13b (n = 11) groups of mice. The bar graphs show the mean ± SEM. *p < 0.05, **p < 0.01 according to Student’s t test. (B) Gross morphology of hearts from 5-month-old WT, R141W, and R141W mice injected with AAV9-PspCas13b-sgRNA2 for 4 weeks. (Scale bars = 500 μm). (C-D) FITC–WGA staining of cardiomyocyte membranes from WT, R141W and R141W + PspCas13b mice (n = 68 cardiomyocytes/group). Scale bar = 100 μm. The bar graphs show the mean ± SEM. ***p < 0.001 according to Student’s t test. (E-I) Left ventricular performance was measured in WT (n = 7), R141W (n = 5) and R141W + PspCas13b (n = 5) groups of mice 4 weeks after the second tail vein injection. EF, left ventricular ejection fraction (F); FS, left ventricular fractional shortening (G); LVID;s, systolic left ventricular internal diameter (H); LVPW;s, systolic left ventricular posterior wall thickness (I). (J, K) Representative Masson staining and quantitation of fibrosis in WT, R141W and R141W + PspCas13b mouse hearts (n = 6 mice/group). Scale bar = 50 μm. (L, M) Quantification of the relative expression of the fibrosis-related genes collagen type I (Col1a1) and periostin (Postn) in hearts from WT, R141W and R141W + PspCas13b mice aged 5 months by qRT-PCR using the 2-∆∆Ct method (n = 3). The loading was normalized to β-actin. (N, O) Quantification of the relative expression of the heart failure genes natriuretic peptide type A (Nppa) and natriuretic peptide type B (Nppb) in hearts from WT, R141W and R141W + PspCas13b mice aged 5 months by qRT-PCR using the 2-∆∆Ct method (n = 3). The loading was normalized to β-actin. The bar graphs show the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 by one-way ANOVA [53,54,34]