EXPERIMENT_TITLE: GSM3898438: Hi-C: SA2-AID clone 1 noauxin; Homo sapiens; Hi-C
EXPERIMENT_DESIGN_LIBRARY_DESCRIPTOR_LIBRARY_CONSTRUCTION_PROTOC: Biotynilated DNA (Hi-C ligation junctions) was isolated using standard phenol/chloroform extraction followed by EtOH precipitation, sonicated to ~800 bp and repurified. Libraries were prepared as described (Zirkel et al, Mol Cell, 2018) on the basis of the Illumina Mate-Pair protocol, by ligating sequencing linkers onto streptavidin-captured biotinylated DNA. All resulting DNA libraries were paired-end sequenced on an Illumina HSeq4000 platform to ~300 million read pairs each on a HiSeq4000 platform (Illumina).
STUDY_DESCRIPTOR_STUDY_ABSTRACT: Cohesin is a ring-shaped multiprotein complex that is crucial for 3D genome organization and transcriptional regulation during differentiation and development. It also confers sister chromatid cohesion and facilitates DNA damage repair. Besides its core subunits SMC3, SMC1A and RAD21, cohesin contains in somatic cells one of two orthologous STAG subunits, SA1 or SA2. How these variable subunits affect the function of the cohesin complex is still unclear. SA1- and SA2-cohesin were initially proposed to organize cohesion at telomeres and centromeres, respectively. Here, we uncover redundant and specific roles of SA1 and SA2 in gene regulation and chromatin looping using HCT116 cells with an auxin-inducible degron (AID) tag fused to either SA1 or SA2. Following rapid depletion of either subunit, we perform high resolution Hi-C, RNA-sequencing and sequential ChIP studies to show that SA1 and SA2 do not co-occupy individual binding sites and have distinct ways how they affect looping and gene expression. These findings are supported at the single cell level by single-molecule localizations via dSTORM super-resolution imaging. Since somatic and congenital mutations of the SA subunits are associated with cancer (SA2) and intellectual disability syndromes with congenital abnormalities (SA1 and SA2), we verified SA1-/SA2-dependencies using human neural stem cells, hence highlighting their importance for understanding particular disease contexts. Overall design: HiC experiments on HCT116 cells with an auxin-inducible degron (AID) tag fused to either the SA1 or SA2 cohesin complex subunit.
SAMPLE_TITLE: Hi-C: SA2-AID clone 1 noauxin
SAMPLE_SAMPLE_ATTRIBUTES_SAMPLE_ATTRIBUTE: [{"TAG": "source_name", "VALUE": "HCT116 cells"}, {"TAG": "cell type", "VALUE": "HCT116 CMV-OsTIR1"}, {"TAG": "genotype", "VALUE": "STAG2-mAID-clover"}, {"TAG": "library id", "VALUE": "79645"}, {"TAG": "treatment", "VALUE": "no auxin"}]
Pool_Member_@attributes_sample_title: Hi-C: SA2-AID clone 1 noauxin
RUN_SET_RUN_Pool_Member_@attributes_sample_title: Hi-C: SA2-AID clone 1 noauxin
RUN_SET_RUN_SRAFiles_SRAFile: [{"Alternatives": [null, null, null], "@attributes": {"cluster": "public", "filename": "SRR9328808.sralite", "url": "https://sra-downloadb.be-md.ncbi.nlm.nih.gov/sos5/sra-pub-zq-14/SRR009/328/SRR9328808.sralite.1", "size": "15037091356", "date": "2020-06-23 19:11:58", "md5": "5eee6fa3b26fa3960a7c9591c669c519", "version": "1", "semantic_name": "SRA Lite", "supertype": "Primary ETL", "sratoolkit": "1"}}, {"Alternatives": null, "@attributes": {"cluster": "public", "filename": "SRR9328808", "url": "https://sra-pub-run-odp.s3.amazonaws.com/sra/SRR9328808/SRR9328808", "size": "24601536928", "date": "2019-06-20 09:05:19", "md5": "c1c4e690708643045f28a6c7e9c58ce7", "version": "1", "semantic_name": "SRA Normalized", "supertype": "Primary ETL", "sratoolkit": "1"}}, {"Alternatives": null, "@attributes": {"cluster": "public", "filename": "SA2-AID_clone1_noaux_rep1_R2.fastq.gz", "size": "20547673814", "date": "2019-06-20 08:44:43", "md5": "03224b08fca6447213ce93aef7c0c158", "version": "1", "semantic_name": "fastq", "supertype": "Original", "sratoolkit": "0"}}, {"Alternatives": null, "@attributes": {"cluster": "public", "filename": "SA2-AID_clone1_noaux_rep1_R1.fastq.gz", "size": "17053260378", "date": "2019-06-20 08:20:56", "md5": "53cbfe997c31d8518dc6bc3038a43692", "version": "1", "semantic_name": "fastq", "supertype": "Original", "sratoolkit": "0"}}]