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Series GSE72631 Query DataSets for GSE72631
Status Public on Jan 12, 2016
Title Effects on the transcriptome upon deletion of distal elements are not correlated with the size of H3K27Ac peaks in human cells
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Genome binding/occupancy profiling by high throughput sequencing
Summary Genome-wide association studies (GWAS) have identified single nucleotide polymorphisms (SNPs) associated with increased risk for colorectal cancer (CRC). A molecular understanding of the functional consequences of this genetic variation is complicated because most GWAS SNPs are located in non-coding regions. After identifying H3K27Ac peaks in HCT116 colon cancer cells that harbor SNPs associated with an increased risk for CRC, we used CRISPR/Cas9 nuclease to delete 2 CRC risk-associated H3K27Ac peaks (E7 and E24), a peak that is not associated with CRC risk (18qE) and a region that doesn?t have H3K27Ac peak (18qNE) from HCT116 cells and analyzed effects on the transcriptome and epigenome. We also deleted E7 region from HEK 293 cells and analyzed effects on the transcriptome of HEK 293. We also confirmed the physical interaction between enhancers of our interest and their putative target genes.
Overall design Analysis of RNA-seq data and ChIP-seq between control clones and enhancer deleted clones in HCT116 cell. For Control, gRNA empty vectorplasmid was transfected with Cas9-GFP. For Deletion, gRNAs that have enhancer target sequences were transfected along with Cas9-GFP. Cells with high GFP expression were identified using fluorescence-activated cell sorting. Sorted cells were plated into individual wells of a 24 well plate and then re-plated as single cells in 10cm dishes and subsequently expanded for further analyses. Samples used in this study were clonal populations. Interaction profiling of enhancers of interest with putative target genes using 4C-seq
Contributor(s) Tak YG
Citation(s) 26743005
Submission date Sep 02, 2015
Last update date May 15, 2019
Contact name Peggy Farnham
Organization name University of Southern California
Department GMCB
Lab NRT6514
Street address 1450 Biggy St.
City LA
State/province CA
ZIP/Postal code 90033
Country USA
Platforms (2)
GPL11154 Illumina HiSeq 2000 (Homo sapiens)
GPL18573 Illumina NextSeq 500 (Homo sapiens)
Samples (32)
GSM1866972 Input_rep1_Control_HCT116_ChIPseq
GSM1866973 Input_rep2_Control_HCT116_ChIPseq
GSM1866974 H3K27Ac_Control_1_USC646_ChIPseq
BioProject PRJNA294738
SRA SRP063339

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Supplementary file Size Download File type/resource
GSE72631_RAW.tar 1.1 Gb (http)(custom) TAR (of TXT, ZIP)
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Raw data are available in SRA
Processed data provided as supplementary file

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