TT-TL-seq reveals transcriptional profiles that accompany DoG induction after hyperosmotic stress.

TT-TimeLapse-seq was used to investigate the nascent transcription profiles that accompany the induction of downstream-of-gene (DoG) containing transcripts in human cell lines. Data from HEK-293T cells exposed to hyperosmotic stress and cells transfected with siRNAs against Integrator subunit 11 are included. Samples were treated with 80 mM KCl for 1 hour or with siRNAs for 48 hours. During the last 5 minutes of treatment, nascent RNAs were labeled with 1 mM s4U. After RNA extractions using TRIzol, 4% RNA from Drosophila S2 cells was added to each sample. TT-TL-seq preparations were done as described in Schofield et al. 2018.

本研究采用TT-TimeLapse-seq技术，探究了人类细胞系中伴随含基因下游（downstream-of-gene，DoG）转录本诱导过程的新生转录谱。实验纳入经高渗胁迫处理的HEK-293T细胞，以及转染靶向整合子复合物亚基11（Integrator subunit 11）小干扰RNA（small interfering RNAs，siRNAs）的细胞。所有样本分别以80 mM氯化钾处理1小时，或用siRNAs处理48小时；在处理的最后5分钟，使用1 mM的s4U对新生RNA进行标记。采用TRIzol试剂提取总RNA后，向每份样本中加入4%的果蝇S2细胞总RNA。TT-TL-seq文库制备流程参照斯科菲尔德等人2018年发表的方法完成。