LINE1 and PRC2 control nucleolar organization and repression of the 8C-state in human ESCs [RNA-Seq_RSeT_H9_L1KD]

To study L1 knockdown effect in RSeT naïve hES cells In this study, we investigated LINE1 (L1), a major family of transposon elements (TEs) in the human genome, in naive hES cells for its roles in regulating 8-cell (8C) stage-specific totipotent genes. RSeT media maintained naive H9 hES cells were transfected with 6-FAM (Fluorescein) labeled non-specific targeting ASO (ASO-Ctr), L1 targeting sense oligo (SO-L1) and anti-sense oligo (ASO-L1), respectively. Cells were maintained in the RSeT media post transfection. 48 hours later, FAM+ cells were FACS collected and immediately processed for RNA extraction and RNA-seq library prep.

本研究旨在探究L1基因敲低在RSeT培养基培养的初始态人类胚胎干细胞（naïve hES cells）中的相关效应。本研究以人类基因组中主要转座元件（transposon elements, TEs）家族之一的LINE1（L1）为研究对象，探究其在该类初始态人类胚胎干细胞中对8细胞（8C）阶段特异性全能基因的调控作用。实验中，将经RSeT培养基维持培养的初始态H9人类胚胎干细胞分别转染6-羧基荧光素（6-FAM，Fluorescein）标记的非特异性靶向反义寡核苷酸（ASO-Ctr）、L1靶向有义寡核苷酸（SO-L1）以及L1靶向反义寡核苷酸（ASO-L1）；转染完成后，细胞仍于RSeT培养基中继续培养。转染48小时后，通过荧光激活细胞分选（FACS）收集FAM阳性细胞，并立即开展RNA提取与RNA测序文库制备工作。