Fig. 6: Disrupting PTBP binding in SYNGAP1 exon 11x upregulates SYNGAP1.
From: Mapping PTBP2 binding in human brain identifies SYNGAP1 as a target for therapeutic splice switching
a Top, Scheme depicting the initial 5-nt resolution ASO walk. The target region spans 93 nt of non-productive 3’ss in SYNGAP1. Bottom, RT-PCR from HEK293T cells transfected with 100 nM of ASO for 24 h, including a positive control ASO targeting SYNGAP1 site 1 (ET-019), a non-targeting ASO control (ET-SC) and no ASO control (Mock, -). b Top, Scheme depicting a combined 2-nt and 1-nt resolution ASO walk. The target region spans 38 nt of non-productive 3’ss in SYNGAP1. Bottom, RT-PCR from HEK293T cells transfected with 100 nM of ASO for 24 h, including the same controls as in a. c qPCR quantification of SYNGAP1 transcript levels from samples in b. d Non-productive and productive transcript levels calculated from densitometric analysis of RT-PCR products from b and represented as log2FC values relative to Mock. Arrows indicate ASOs that increase SYNGAP1 productive transcript and mRNA levels. Black line indicates ASOs that lead to no-go decay. e Top and left panel: RT-PCR from HEK293T cells transfected with increasing concentrations of STK-071, ET-085, and ET-SC for 48 h. Right panel: qPCR showing SYNGAP1 mRNA levels. f Western blot from HEK293T cells transfected as in e. Data are represented as mean values ± SEM. All data points represent independent biological replicates. a–d (n = 2). e, f (n = 3). e, f One-way ANOVA with Dunnett’s multiple comparison test vs. mock-treated cells (–). Source data are provided as a Source Data file. FC fold change.
