LINE1 and PRC2 control nucleolar organization and repression of the 8C-state in human ESCs [RNA-Seq_RSeT+DT_H9_L1KD+siRNA-KD]

To study siRNA-mediated TPRX1, TP53 and H3.XY co-knockdown effected when transfected together with L1 targeting anti sense oligos (ASO-L1) in RSeT+DT naïve hES cells. In this study, we investigated if co-knockdown of TPRX1, TP53 or H3.XY expression would reverse the ASO-L1 knockdown induced 8-cell (8C) stage-specific genes upregulation. RSeT media maintained naive H9 hES cells were transfected with non-specific targeting control anti-sense oligos (ASO-Ctr) or ASO-L1, together with 6-FAM (Fluorescein) labeled siRNAs targeting non-specific sequence (NTsi), TPRX1 (TPRX1si), TP53 (TP53si) and H3.XY (H3.XYsi), respectively. Transfected cells were maintained in the RSeT media added with 10 nM DZNep and 5nM TSA (RSeT+DT). 48 hours later, FAM+ cells were FACS collected and immediately processed for RNA extraction and RNA-seq library prep.