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Series GSE117144 Query DataSets for GSE117144
Status Public on Oct 01, 2018
Title Comparison of CWC27 knockdown in HEK293T cells compared to scrambled shRNA control
Organism Homo sapiens
Experiment type Expression profiling by array
Summary HEK293T cells transduced with shRNA from MISSION library TRCN0000000139 using lentiviral delivery system. HEK293T cells transduced with scrambled shRNA, gifted from Dr. Mauricio Reginato. Sequence is 5′-CCTAAGGTTAAGTCGCCCTCGCTCTAGCGAGGGCGACTTAACCTT-3′. Tara L Davis, S. RaElle Jackson, Beth Adams, Anh Trinh, Peter Naranjo, and Angie Giang performed primary experimental contributions to cell lines, RNA/cDNA preparation, and validation of results, all Drexel University College of Medicine, Philadelphia, PA 19102. Hetty Rodriguez and John Tobias performed Bioanalyzer and microarray expreriments, and initial data processing. Affiliation: Molecular Profiling Facility and Genomic Analysis Core Bioinformatics Group, University of Pennsylvania.
Human CWC27 (aliases: NY-CO-10, Serologically Defined Colon Cancer Antigen 10 or SDCCAG-10) is a cyclophilin, an enzyme that interconverts cis and trans isomers of proline. The CWC27 gene, in addition to the cyclophilin domain, encodes for two low complexity regions and a coiled coil region. CWC27 associates with the human spliceosome, the complex and dynamic machinery that removes intronic sequence from pre-messenger RNA (pre-mRNA). Nothing is known about the function of CWC27 in the nucleus. To understand the function of CWC27, we knocked down CWC27 in human cells. We characterized a set of alternative splicing and transcriptional events that are CWC27-responsive. We used these splicing and transcriptional bioassays to show that CWC27-responsive events are largely specific, even within the cyclophilin family.
Overall design 3 replicates of CWC27 knockdown cell lines (HEK293T cells, stably integrated, selected using puro, MISSION shRNA TRCN0000000139: 5'-CCGGACGGGAACTCTTAGCAGCAAACTCGAGTTTGCTGCTAAGAGTTCCCGTTTTTT-3'). 3 replicates of SCR control cell lines (HEK293T cells, stably integrated, selected using puro, sequence 5′-CCTAAGGTTAAGTCGCCCTCGCTCTAGCGAGGGCGACTTAACCTT-3′). total RNA purified from ~1 million cells, cDNA converted using random hexamer primers, quantity measured using NanoDrop, quality quantified using BioAnaylzer. Affymatrix HTA2.0 array
Contributor(s) Davis TL, Jackson SR, Adams B, Trinh A, Naranjo P, Giang A, Rodriguez H, Tobias J
Citation(s) 30518120
NIH grant(s)
Grant ID Grant title Affiliation Name
R00 GM094293 Functional and structural characterization of spliceosomal cyclophilins DREXEL UNIVERSITY Tara L Davis
Submission date Jul 16, 2018
Last update date May 15, 2019
Contact name Tara Davis
Organization name Drexel University College of Medicine
Department Biochemistry and Molecular Biology
Lab Lab 10127
Street address 245 N. 15th St. MS 497
City Philadelphia
State/province PA
ZIP/Postal code 19102
Country USA
Platforms (2)
GPL17585 [HTA-2_0] Affymetrix Human Transcriptome Array 2.0 [probe set (exon) version]
GPL17586 [HTA-2_0] Affymetrix Human Transcriptome Array 2.0 [transcript (gene) version]
Samples (12)
GSM3272154 HEK293T_139CWC27_gene_rep1
GSM3272155 HEK293T_139CWC27_gene_rep2
GSM3272156 HEK293T_139CWC27_gene_rep3
BioProject PRJNA481269

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE117144_RAW.tar 383.7 Mb (http)(custom) TAR (of CEL, CHP)
Processed data included within Sample table
Processed data provided as supplementary file

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