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E-MTAB-6564

ABSTRACT: RNA-Seq of control, RNPS1 knockdown and rescue in HeLa cells

PROVIDER: E-MTAB-6564 | ExpressionAtlas | 2025-01-23

REPOSITORIES: ExpressionAtlas

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			Action	DRS
	E-MTAB-6564-analysis-methods.tsv	Tabular
	E-MTAB-6564-analytics.tsv	Tabular
	E-MTAB-6564-atlasExperimentSummary.Rdata	Rdata
	E-MTAB-6564-configuration.xml	Xml
	E-MTAB-6564-g4_g1-mvaPlot.png	Other

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Project description: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.

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Project description:We report the knockdown and rescue of RNPS1, a component of the ASAP and PSAP complexes and, hence, an auxiliary component of the exon junction complex. The studied conditions show an extensive change in alternative splicing and the cell's transcriptome. For the knockdown, HeLa cells were transfected with siRNAs targeting RNPS1 or the Luciferase control. Total RNA was extracted with peqGOLD TriFast, underwent ribosomal depletion and strand-specific library preparation (TruSeq R Stranded Total RNA LT). Sequencing occurred in an Illumina HiSeq4000 sequencer with 2×75bp, producing roughly 35 million read-pair per sample.

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