Characterization of E2F8-mediated transcriptional regulation in lethal prostate cancer
To identify the genomic bindings and transcriptional targets of E2F8 in lethal prostate cancer cells. To examine the potential off-target effects by CRISPR/CasRx-mediated E2F8 knockdown in 293FT cells. To analyze differentially expressed genes upon TT3 LLN-CRISPR/CasRx-mediated E2F8 knockdown in DU145 xenografts. Overall design: We performed E2F8 ChIP-seq to map genome-wide E2F8 binding sites in PC3 and DU145 cells. To identify E2F8-regulated genes, RNA-seq was performed in PC3 and DU145 cells transfected with control siRNA or siRNA targeting E2F8 for 72h. To evaluate the off-target effects of CasRx-mediated E2F8 silencing, RNA-seq was conducted in 293FT cells transfected with either a CasRx expression plasmid along with a non-targeting pre-gRNA control(pre-gCtrl), or a pre-gRNA targeting E2F8(pre-gE2F8) plasmid for 24h. To elucidate the molecular mechanisms by which TT3 lipid-like nanoparticles(LLNs) carrying CasRx/pre-gE2F8 inhibit DU145 xenografts growth, RNA-seq was performed to identify the differentially expressed genes between TT3 LLN-CasRx/pre-gCtrl and TT3 LLN-CasRx/pre-gE2F8 treated groups.