Extended Data Fig. 2: gRNA screening and comparison of EpiReg-C and EpiReg-T.
a, b, Bar graph showing the effect of different gRNAs on PCSK9 repression using the V1 version of EpiReg mRNA in Huh7 cells (a) and primary monkey hepatocytes (b). A combination of Hs.gRNA1 + 2 (a) or Mfa.gRNA 5 + 12 (b) was selected for use in subsequent experiments. PCSK9 expression was measured by qPCR 7 days post-transfection. Data are presented as mean ± SEM, n = 3 independent experiments per group. c, The comparison between EpiReg-C and EpiReg-T at high doses in the second batch of humanized PCSK9 mice. Bar graph comparing the efficiency of PCSK9 repression. EpiRegs were administered via tail vein injection, and samples were collected 21 days post-injection. PCSK9 levels in the serum were measured by ELISA. Data are presented as mean ± SEM, n = 5 animals per group. To compare the differences, a two-tailed, unpaired Mann-Whitney test was performed. ns, not significant. d-f, Dose-response curves comparing the efficiency of EpiReg-C and EpiReg-T in Huh7 cells (d), PHH (e), and humanized PCSK9 mice (f). The half-maximum effective concentration (EC50) and the coefficient of determination (R2) for each group are indicated, calculated by fitting a four-parameter logistic model (R2 > 0.8 for all treatments). Dashed lines represent 50% inhibition. Data are presented as mean ± SEM, n = 3 for Huh7 and PHH, n = 5 for humanized PCSK9 mice. g, Bar graph comparing the EC50 values of EpiReg-C and EpiReg-T Huh7 cells, PHH, and humanized PCSK9 mice.
