RNA-Seq of control and RNPS1 knockdown with or without RNPS1-RRM domain rescue in HeLa Tet-Off cells. RNA-Seq of control and RNPS1 knockdown with or without RNPS1-RRM domain rescue in HeLa Tet-Off cells

RNPS1 is a splicing regulatory protein and a component of the ASAP/PSAP complex, which is associated with the exon junction complex and modulates alternative splicing. It was previously postulated that the isolated RRM domain of RNPS1 in complex with ASAP/PSAP is able to regulate certain alternative splicing events. We aimed to investigate in HeLa Tet-Off cells which alternative splicing events are rescued by the expression of the isolated RRM domain of RNPS1 in a RNPS1 knockdown background by using RNA-Seq analyses. The rescue construct was stably integrated into the genome using the PiggyBac transposon system. As controls, either Luciferase (Luc) siRNA was used or RNPS1 was knocked down without rescue.

RNPS1是一种剪接调控蛋白，同时也是ASAP/PSAP复合物（ASAP/PSAP complex）的组成组分，该复合物可与外显子连接复合物（exon junction complex）相结合，并调控可变剪接（alternative splicing）过程。此前曾有假说提出，RNPS1的独立RRM结构域（RNA recognition motif domain）与ASAP/PSAP复合物结合后，能够调控特定的可变剪接事件。本研究旨在通过RNA测序（RNA-Seq）分析，在HeLa Tet-Off细胞（HeLa Tet-Off cells）中探究：在RNPS1敲低的细胞背景下，过表达RNPS1的独立RRM结构域可挽救哪些可变剪接事件。本研究中的挽救型重组载体通过PiggyBac转座子系统（PiggyBac transposon system）稳定整合至宿主基因组。实验设置两组对照：一组采用荧光素酶（Luciferase）小干扰RNA（siRNA）处理，另一组则仅进行RNPS1敲低而不施加挽救操作。