Fig. 3: In vivo regulation of an AAV-expressed reporter transgene by exogenous delivery of siRNA and REVERSIR.

a Diagram depicting dual agent approach involving suppression of AAV transgene expression with siRNA and subsequent upregulation of protein expression by abrogation of siRNA activity with REVERSIR. b Mice were injected with 2 × 10¹¹ GC of AAV encoding bicistronic expression of a GLuc reporter with a TTR siRNA binding site in the 3’ UTR. Mice were subcutaneously injected with vehicle or TTR siRNA at 9 mg/kg (D0). This was followed on D14 with a single molar equivalent dose of full-length 22-mer (3 mg/kg) or 9-mer TTR REVERSIR (1.6 mg/kg) and compared to NT REVERSIR control. c Serum GLuc levels normalized to pre-dose (D0) for each animal (n = 5 for PBS; n = 4 for vehicle and 22-mer TTR REVERSIR groups; n = 3 for 22-mer NT REVERSIR and both 9-mer REVERSIR groups). Data were analyzed by two-way repeated measures ANOVA followed by Tukey’s post hoc test. d Gluc transcript levels in liver tissue at D42 normalized to Gapdh control. e Serum GLuc levels at D21 in AAV-transduced mice that received TTR siRNA (9 mg/kg; D0), followed by increasing doses of 9-mer TTR REVERSIR (D14) or NT REVERSIR (n = 4 for PBS, TTR siRNA, 0.3 mg/kg REVERSIR, and NT REVERSIR groups; n = 5 for 0.5 mg/kg REVERSIR group; n = 3 for 1.6 mg/kg TTR REVERSIR group). Data were analyzed by one-way ANOVA followed by Holm-Sidak or Dunnett’s tests, respectively for (d) and (e). ^*p < 0.05 ^**p < 0.01 ^***p < 0.001 n.s., not significant. Error bars represent s.e.m. Source data are provided as a Source Data file. Diagrams were created with BioRender.com.